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Id Subject Object Predicate Lexical cue
T22839 2636-2637 . denotes .
T22894 2806-2810 JJ denotes wild
T22893 2803-2805 NN denotes bp
T22892 2799-2802 CD denotes 429
T22891 2816-2822 NN denotes signal
T22890 2795-2798 DT denotes the
T22889 2792-2794 IN denotes of
T22888 2784-2791 NN denotes absence
T22887 2780-2783 DT denotes the
T22886 2777-2779 IN denotes by
T22885 2770-2776 VBN denotes marked
T22884 2768-2770 , denotes ,
T22883 2767-2768 SYM denotes -
T22882 2766-2767 HYPH denotes /
T22881 2765-2766 SYM denotes -
T22880 2760-2765 NN denotes Capn2
T22879 2754-2755 CD denotes 4
T22878 2753-2754 SYM denotes #
T22877 2746-2752 NN denotes embryo
T22876 2756-2759 VBD denotes was
T22875 2740-2745 IN denotes while
T22874 2738-2739 SYM denotes -
T22873 2737-2738 HYPH denotes /
T22872 2736-2737 SYM denotes +
T22871 2731-2736 NN denotes Capn2
T22870 2724-2725 CD denotes 6
T22869 2720-2723 CC denotes and
T22868 2718-2720 , denotes ,
T22867 2717-2718 CD denotes 5
T22866 2715-2717 , denotes ,
T22865 2714-2715 CD denotes 3
T22864 2712-2714 , denotes ,
T22863 2711-2712 CD denotes 2
T22862 2709-2711 , denotes ,
T22861 2726-2730 VBD denotes were
T22860 2707-2708 SYM denotes #
T22859 2708-2709 CD denotes 1
T22858 2699-2706 NNS denotes Embryos
T22857 2698-2823 sentence denotes Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal.
T22856 2697-2698 . denotes .
T22855 2689-2691 VBZ denotes is
T22854 2681-2688 NNS denotes embryos
T22853 2675-2676 HYPH denotes -
T22852 2676-2680 NN denotes cell
T22851 2674-2675 CD denotes 8
T22850 2671-2673 IN denotes of
T22849 2660-2670 NN denotes genotyping
T22848 2656-2659 DT denotes the
T22847 2653-2655 IN denotes of
T22846 2645-2652 NN denotes example
T22845 2642-2644 DT denotes An
T22844 2640-2641 -RRB- denotes )
T22843 2692-2697 VBN denotes shown
T22842 2639-2640 LS denotes B
T22841 2638-2639 -LRB- denotes (
T22840 2637-2698 sentence denotes (B) An example of the genotyping of 8-cell embryos is shown.
T22838 2623-2629 NN denotes mutant
T22837 2620-2622 NN denotes bp
T22836 2616-2619 CD denotes 389
T22835 2630-2636 NN denotes signal
T22834 2612-2615 DT denotes the
T22833 2609-2611 IN denotes of
T22832 2601-2608 NN denotes absence
T22831 2597-2600 DT denotes the
T22830 2594-2596 IN denotes by
T22829 2586-2593 VBN denotes denoted
T22828 2584-2586 , denotes ,
T22827 2583-2584 SYM denotes +
T22826 2582-2583 HYPH denotes /
T22825 2581-2582 SYM denotes +
T22824 2576-2581 NN denotes Capn2
T22823 2569-2570 CD denotes 7
T22822 2568-2569 SYM denotes #
T22821 2564-2567 CC denotes and
T22820 2561-2562 SYM denotes #
T22819 2562-2563 CD denotes 3
T22818 2553-2560 NNS denotes embryos
T22817 2571-2575 VBD denotes were
T22816 2545-2552 IN denotes whereas
T22815 2543-2544 SYM denotes -
T22814 2542-2543 HYPH denotes /
T22813 2541-2542 SYM denotes +
T22812 2536-2541 NN denotes Capn2
T22811 2529-2530 CD denotes 6
T22810 2525-2528 CC denotes and
T22809 2523-2525 , denotes ,
T22808 2522-2523 CD denotes 5
T22807 2520-2522 , denotes ,
T22806 2519-2520 CD denotes 4
T22805 2517-2519 , denotes ,
T22804 2516-2517 CD denotes 2
T22803 2514-2516 , denotes ,
T22802 2531-2535 VBD denotes were
T22801 2512-2513 SYM denotes #
T22800 2513-2514 CD denotes 1
T22799 2504-2511 NNS denotes Embryos
T22798 2503-2637 sentence denotes Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal.
T22797 2502-2503 . denotes .
T22796 2489-2494 NN denotes stage
T22795 2495-2502 NNS denotes embryos
T22794 2478-2488 NN denotes blastocyst
T22793 2475-2477 IN denotes of
T22792 2464-2474 NN denotes genotyping
T22791 2460-2463 DT denotes the
T22790 2457-2459 IN denotes of
T22789 2434-2448 JJ denotes representative
T22788 2432-2433 DT denotes A
T22787 2430-2431 -RRB- denotes )
T22786 2449-2456 NN denotes example
T22785 2429-2430 LS denotes A
T22784 2428-2429 -LRB- denotes (
T22783 2427-2503 sentence denotes (A) A representative example of the genotyping of blastocyst stage embryos.
T22782 2426-2427 . denotes .
T22781 2410-2418 JJ denotes internal
T22780 2419-2426 NN denotes control
T22779 2406-2409 DT denotes the
T22778 2402-2405 IN denotes for
T22777 2399-2401 NN denotes bp
T22776 2395-2398 CD denotes 213
T22775 2391-2394 CC denotes and
T22774 2389-2391 , denotes ,
T22773 2376-2382 NN denotes mutant
T22772 2383-2389 NN denotes allele
T22771 2372-2375 DT denotes the
T22770 2368-2371 IN denotes for
T22769 2365-2367 NN denotes bp
T22768 2361-2364 CD denotes 389
T22767 2359-2361 , denotes ,
T22766 2347-2348 HYPH denotes -
T22765 2348-2352 NN denotes type
T22764 2343-2347 JJ denotes wild
T22763 2353-2359 NN denotes allele
T22762 2339-2342 DT denotes the
T22761 2335-2338 IN denotes for
T22760 2332-2334 NN denotes bp
T22759 2328-2331 CD denotes 429
T22758 2323-2327 VBD denotes were
T22757 2308-2313 JJ denotes final
T22756 2314-2322 NNS denotes products
T22755 2304-2307 DT denotes The
T22754 2303-2427 sentence denotes The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control.
T22753 2302-2303 . denotes .
T22752 2295-2302 NNS denotes alleles
T22751 2290-2294 DT denotes both
T22750 2287-2289 IN denotes in
T22749 2278-2280 VBZ denotes is
T22748 2281-2286 VBN denotes found
T22747 2272-2277 WDT denotes which
T22746 2270-2272 , denotes ,
T22745 2254-2263 NN denotes targeting
T22744 2264-2270 NN denotes vector
T22743 2250-2253 DT denotes the
T22742 2247-2249 IN denotes of
T22741 2241-2242 -RRB- denotes )
T22740 2233-2241 JJ denotes upstream
T22739 2232-2233 -LRB- denotes (
T22738 2226-2231 JJ denotes short
T22737 2243-2246 NN denotes arm
T22736 2222-2225 DT denotes the
T22735 2219-2221 IN denotes in
T22734 2211-2218 VBN denotes located
T22733 2209-2211 , denotes ,
T22732 2193-2200 NN denotes control
T22731 2184-2192 JJ denotes internal
T22730 2201-2209 NN denotes sequence
T22729 2181-2183 DT denotes an
T22728 2176-2180 IN denotes with
T22727 2158-2162 VBD denotes were
T22726 2150-2157 NNS denotes alleles
T22725 2143-2149 NN denotes mutant
T22724 2140-2142 CC denotes or
T22723 2134-2135 HYPH denotes -
T22722 2130-2134 JJ denotes wild
T22721 2126-2129 DT denotes the
T22720 2135-2139 NN denotes type
T22719 2119-2125 CC denotes either
T22718 2116-2118 IN denotes in
T22717 2104-2115 RB denotes exclusively
T22716 2098-2103 VBN denotes found
T22715 2079-2088 NNS denotes reactions
T22714 2089-2097 NNS denotes segments
T22713 2070-2078 JJ denotes separate
T22712 2163-2175 VBN denotes co-amplified
T22711 2067-2069 IN denotes In
T22710 2066-2303 sentence denotes In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles.
T22709 2065-2066 . denotes .
T22708 2064-2065 NN denotes K
T22707 2053-2063 NN denotes proteinase
T22706 2048-2052 IN denotes with
T22705 2039-2047 VBN denotes digested
T22704 2034-2038 RB denotes then
T22703 2030-2033 CC denotes and
T22702 2021-2029 NNS denotes oviducts
T22701 2017-2020 DT denotes the
T22700 2012-2016 IN denotes from
T22699 1999-2003 VBD denotes were
T22698 1997-1998 -RRB- denotes )
T22697 1993-1997 NN denotes E2.5
T22696 1992-1993 -LRB- denotes (
T22695 2004-2011 VBN denotes flushed
T22694 1978-1979 HYPH denotes -
T22693 1979-1983 NN denotes cell
T22692 1977-1978 CD denotes 8
T22691 1974-1976 CC denotes or
T22690 1972-1973 -RRB- denotes )
T22689 1968-1972 NN denotes E3.5
T22688 1967-1968 -LRB- denotes (
T22687 1984-1991 NNS denotes embryos
T22686 1956-1966 NN denotes Blastocyst
T22685 1955-2066 sentence denotes Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K.
T22684 1954-1955 . denotes .
T22683 1949-1954 NNS denotes plugs
T22665 1853-1858 NN denotes Capn2
T22664 1852-1955 sentence denotes Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs.
T22663 1851-1852 . denotes .
T22662 1850-1851 CD denotes 4
T22661 1843-1849 NN denotes Figure
T22660 1840-1842 IN denotes in
T22659 1830-1839 VBN denotes described
T22658 1827-1829 IN denotes as
T22657 1814-1826 NN denotes implantation
T22656 1811-1813 IN denotes to
T22655 1805-1810 JJ denotes prior
T22654 1797-1804 NNS denotes embryos
T22653 1788-1796 VB denotes genotype
T22652 1785-1787 TO denotes to
T22651 1776-1779 VBD denotes was
T22650 1780-1784 VBN denotes used
T22649 1763-1766 NN denotes PCR
T22648 1756-1762 VBN denotes nested
T22647 1767-1775 NN denotes strategy
T22646 1754-1755 DT denotes A
T22645 1753-1852 sentence denotes A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4.
T22644 1752-1753 . denotes .
T22643 1745-1752 NNS denotes embryos
T22642 1728-1744 JJ denotes pre-implantation
T22641 1725-1727 IN denotes of
T22640 1714-1724 NN denotes Genotyping
T22907 2863-2864 . denotes .
T22906 2850-2856 NN denotes weight
T22905 2840-2849 JJ denotes molecular
T22904 2857-2863 NN denotes marker
T22903 2836-2839 DT denotes the
T22902 2826-2827 -RRB- denotes )
T22901 2828-2835 VBZ denotes denotes
T22900 2825-2826 NN denotes M
T22899 2824-2825 -LRB- denotes (
T22682 1941-1948 JJ denotes vaginal
T22681 1938-1940 IN denotes of
T22680 1927-1937 NN denotes appearance
T22679 1923-1926 DT denotes the
T22678 1920-1922 IN denotes by
T22677 1894-1907 NN denotes fertilization
T22676 1891-1893 IN denotes of
T22675 1908-1919 VBN denotes established
T22674 1886-1890 NN denotes date
T22673 1882-1885 DT denotes the
T22672 1878-1881 CC denotes and
T22671 1867-1871 VBD denotes were
T22670 1872-1877 VBN denotes mated
T22669 1860-1861 SYM denotes -
T22668 1859-1860 HYPH denotes /
T22667 1858-1859 SYM denotes +
T22666 1862-1866 NNS denotes mice
T22084 1702-1703 . denotes .
T22083 1689-1695 NN denotes mutant
T22082 1696-1702 NN denotes allele
T22081 1685-1688 DT denotes the
T22080 1682-1684 IN denotes in
T22079 1668-1669 HYPH denotes -
T22078 1669-1672 NN denotes Neo
T22077 1665-1668 NN denotes PGK
T22076 1673-1681 NN denotes cassette
T22075 1661-1664 DT denotes the
T22074 1658-1660 IN denotes by
T22073 1646-1648 VBZ denotes is
T22072 1649-1657 VBN denotes replaced
T22071 1641-1645 WDT denotes that
T22070 1628-1629 HYPH denotes -
T22069 1629-1633 NN denotes type
T22068 1624-1628 JJ denotes wild
T22067 1634-1640 NN denotes allele
T22066 1620-1623 DT denotes the
T22065 1617-1619 IN denotes of
T22064 1609-1616 NN denotes segment
T22063 1605-1608 DT denotes the
T22062 1594-1604 VBZ denotes delineates
T22061 1573-1583 JJ denotes horizontal
T22060 1571-1573 , denotes ,
T22059 1567-1571 JJ denotes grey
T22058 1584-1593 NN denotes rectangle
T22057 1565-1566 DT denotes A
T22056 1564-1703 sentence denotes A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele.
T22055 1563-1564 . denotes .
T22054 1558-1559 -RRB- denotes )
T22053 1548-1558 JJ denotes downstream
T22052 1547-1548 -LRB- denotes (
T22051 1542-1546 JJ denotes long
T22050 1560-1563 NN denotes arm
T22049 1538-1541 DT denotes the
T22048 1535-1537 IN denotes of
T22047 1529-1530 SYM denotes '
T22046 1528-1529 CD denotes 5
T22045 1531-1534 NN denotes end
T22044 1524-1527 DT denotes the
T22043 1520-1523 CC denotes and
T22042 1510-1515 JJ denotes short
T22041 1516-1519 NN denotes arm
T22040 1506-1509 DT denotes the
T22039 1503-1505 IN denotes of
T22038 1492-1502 NNS denotes boundaries
T22037 1488-1491 DT denotes the
T22036 1466-1474 JJ denotes vertical
T22035 1475-1480 NNS denotes lines
T22034 1461-1465 JJ denotes thin
T22033 1481-1487 VBP denotes denote
T22032 1455-1460 IN denotes while
T22031 1453-1455 , denotes ,
T22030 1433-1443 JJ denotes horizontal
T22029 1431-1433 , denotes ,
T22028 1427-1431 JJ denotes open
T22027 1444-1453 NN denotes rectangle
T22026 1424-1426 DT denotes an
T22025 1421-1423 IN denotes by
T22024 1407-1408 HYPH denotes -
T22023 1408-1411 NN denotes Neo
T22022 1404-1407 NN denotes PGK
T22021 1400-1403 DT denotes the
T22020 1398-1400 , denotes ,
T22019 1379-1387 JJ denotes vertical
T22018 1377-1379 , denotes ,
T22017 1388-1398 NNS denotes rectangles
T22016 1373-1377 JJ denotes open
T22015 1370-1372 IN denotes by
T22014 1412-1420 NN denotes cassette
T22013 1354-1357 VBP denotes are
T22012 1358-1369 VBN denotes represented
T22011 1348-1353 NNS denotes Exons
T22010 1347-1564 sentence denotes Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm.
T22009 1346-1347 . denotes .
T22008 1331-1336 CD denotes three
T22007 1337-1346 NNS denotes reactions
T22006 1327-1330 DT denotes the
T22005 1324-1326 IN denotes of
T22004 1319-1323 DT denotes each
T22003 1315-1318 IN denotes for
T22002 1308-1309 -RRB- denotes )
T22001 1302-1308 VBN denotes nested
T22000 1301-1302 -LRB- denotes (
T21999 1310-1314 NNS denotes sets
T21998 1297-1300 CD denotes two
T21997 1294-1296 IN denotes in
T21996 1281-1284 VBP denotes are
T21995 1279-1281 , denotes ,
T21994 1270-1279 NNS denotes triangles
T21993 1267-1269 IN denotes by
T21992 1255-1266 VBN denotes represented
T21991 1253-1255 , denotes ,
T21990 1285-1293 VBN denotes depicted
T21989 1246-1253 NNS denotes primers
T21988 1242-1245 DT denotes The
T21987 1241-1347 sentence denotes The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions.
T21986 1240-1241 . denotes .
T21985 1231-1232 HYPH denotes -
T21984 1232-1240 JJ denotes specific
T21983 1225-1231 NN denotes allele
T21982 1202-1211 NN denotes antisense
T21981 1212-1219 NNS denotes primers
T21980 1198-1201 DT denotes the
T21979 1220-1224 VBD denotes were
T21978 1190-1197 IN denotes whereas
T21977 1188-1190 , denotes ,
T21976 1179-1188 NNS denotes reactions
T21975 1174-1178 DT denotes both
T21974 1171-1173 IN denotes in
T21973 1159-1160 '' denotes '
T21972 1161-1170 NNS denotes sequences
T21971 1149-1159 JJ denotes diagnostic
T21970 1148-1149 `` denotes '
T21969 1140-1147 VB denotes amplify
T21968 1137-1139 TO denotes to
T21967 1127-1131 VBD denotes were
T21966 1132-1136 VBN denotes used
T21965 1113-1118 NN denotes sense
T21964 1108-1112 JJ denotes same
T21963 1119-1126 NNS denotes primers
T21962 1104-1107 DT denotes The
T21961 1103-1241 sentence denotes The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific.
T21960 1102-1103 . denotes .
T21959 1086-1094 JJ denotes internal
T21958 1095-1102 NN denotes control
T21957 1083-1085 DT denotes an
T21956 1080-1082 IN denotes as
T21955 1069-1070 '' denotes '
T21954 1059-1069 JJ denotes diagnostic
T21953 1058-1059 `` denotes '
T21952 1071-1079 NNS denotes products
T21951 1054-1057 DT denotes the
T21950 1049-1053 IN denotes with
T21949 1032-1035 VBD denotes was
T21948 1022-1027 JJ denotes short
T21947 1028-1031 NN denotes arm
T21946 1018-1021 DT denotes the
T21945 1011-1017 IN denotes within
T21944 1003-1010 VBN denotes located
T21943 991-993 NN denotes bp
T21942 987-990 CD denotes 213
T21941 994-1002 NN denotes sequence
T21940 985-986 DT denotes a
T21939 983-985 , denotes ,
T21938 974-983 NNS denotes reactions
T21937 969-973 DT denotes both
T21936 1036-1048 VBN denotes co-amplified
T21935 966-968 IN denotes In
T21934 965-1103 sentence denotes In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control.
T21933 964-965 . denotes .
T21932 948-957 NN denotes targeting
T21931 958-964 NN denotes vector
T21930 944-947 DT denotes the
T21929 941-943 IN denotes of
T21928 935-936 -RRB- denotes )
T21927 927-935 JJ denotes upstream
T21926 926-927 -LRB- denotes (
T21925 920-925 JJ denotes short
T21924 937-940 NN denotes arm
T21923 916-919 DT denotes the
T21922 913-915 IN denotes of
T21921 907-908 SYM denotes '
T21920 906-907 CD denotes 3
T21919 909-912 NN denotes end
T21918 902-905 DT denotes the
T21917 893-901 VBG denotes spanning
T21916 888-892 DT denotes both
T21915 886-888 , denotes ,
T21914 873-879 NN denotes mutant
T21913 880-886 NN denotes allele
T21912 869-872 DT denotes the
T21911 864-868 IN denotes from
T21910 853-855 NN denotes bp
T21909 849-852 CD denotes 389
T21908 856-863 NN denotes segment
T21907 847-848 DT denotes a
T21906 843-846 CC denotes and
T21905 830-831 HYPH denotes -
T21904 831-835 NN denotes type
T21903 826-830 JJ denotes wild
T21902 836-842 NN denotes allele
T21901 822-825 DT denotes the
T21900 817-821 IN denotes from
T21899 805-807 NN denotes bp
T21898 801-804 CD denotes 429
T21897 808-816 NN denotes fragment
T21896 799-800 DT denotes a
T21895 791-798 VB denotes amplify
T21894 788-790 TO denotes to
T21893 778-782 VBD denotes were
T21892 783-787 VBN denotes used
T21891 768-777 NNS denotes reactions
T21890 759-767 JJ denotes Separate
T21889 758-965 sentence denotes Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector.
T21888 757-758 . denotes .
T21887 750-757 NNS denotes embryos
T21886 733-749 JJ denotes pre-implantation
T21885 724-732 VB denotes genotype
T21884 721-723 TO denotes to
T21883 715-720 NN denotes order
T21882 712-714 IN denotes in
T21881 698-701 VBD denotes was
T21880 685-688 NN denotes PCR
T21879 678-684 VBN denotes nested
T21878 689-697 NN denotes strategy
T21877 676-677 DT denotes a
T21876 674-676 , denotes ,
T21875 658-665 JJ denotes genetic
T21874 666-674 NN denotes material
T21873 646-657 JJ denotes extractable
T21872 643-645 IN denotes of
T21871 634-642 NN denotes scarcity
T21870 630-633 DT denotes the
T21869 627-629 IN denotes to
T21868 702-711 VBN denotes developed
T21867 623-626 IN denotes Due
T21866 622-758 sentence denotes Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos.
T21865 621-622 . denotes .
T21864 614-621 NNS denotes embryos
T21863 597-613 JJ denotes pre-implantation
T21862 594-596 IN denotes of
T21861 583-593 NN denotes genotyping
T21860 579-582 IN denotes for
T21859 566-569 NN denotes PCR
T21858 570-578 NN denotes strategy
T21857 559-565 VBN denotes Nested
T7021 2971-2972 . denotes .
T7020 2970-2971 CD denotes 2
T7019 2964-2969 NN denotes Table
T7018 2961-2963 IN denotes in
T7017 2951-2954 VBP denotes are
T22898 2823-2864 sentence denotes (M) denotes the molecular weight marker.
T22897 2822-2823 . denotes .
T22896 2810-2811 HYPH denotes -
T22895 2811-2815 NN denotes type
T22936 10-18 NN denotes Genotype
T22937 19-31 NN denotes distribution
T22938 32-34 IN denotes of
T22939 35-44 NN denotes offspring
T22940 45-52 VBN denotes derived
T22941 53-57 IN denotes from
T22942 58-63 NN denotes Capn2
T22943 75-79 NNS denotes mice
T22944 64-74 JJ denotes transgenic
T22945 79-80 . denotes .
T7016 2926-2942 JJ denotes pre-implantation
T7015 2922-2925 CC denotes and
T7014 2920-2922 , denotes ,
T7013 2943-2950 NNS denotes embryos
T7012 2903-2920 JJ denotes post-implantation
T7011 2901-2903 , denotes ,
T7010 2892-2901 NNS denotes weanlings
T7009 2888-2891 IN denotes for
T7008 2880-2887 VBZ denotes results
T7007 2955-2960 VBN denotes shown
T7006 2869-2879 NN denotes genotyping
T7005 1-2868 DT denotes Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The
T7004 0-2972 sentence denotes Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The genotyping results for weanlings, post-implantation, and pre-implantation embryos are shown in Table 2.
R1892 T7017 T7007 auxpass are,shown
R1893 T7018 T7007 prep in,shown
R1894 T7019 T7018 pobj Table,in
R1895 T7020 T7019 nummod 2,Table
R1896 T7021 T7007 punct .,shown
R6470 T21857 T21858 amod Nested,strategy
R6471 T21859 T21858 compound PCR,strategy
R6472 T21860 T21858 prep for,strategy
R6473 T21861 T21860 pobj genotyping,for
R6474 T21862 T21861 prep of,genotyping
R6475 T21863 T21864 amod pre-implantation,embryos
R6476 T21864 T21862 pobj embryos,of
R6477 T21865 T21858 punct .,strategy
R6478 T21867 T21868 prep Due,developed
R6479 T21869 T21867 pcomp to,Due
R6480 T21870 T21871 det the,scarcity
R6481 T21871 T21867 pobj scarcity,Due
R6482 T21872 T21871 prep of,scarcity
R6483 T21873 T21874 amod extractable,material
R6484 T21874 T21872 pobj material,of
R6485 T21875 T21874 amod genetic,material
R6486 T21876 T21868 punct ", ",developed
R6487 T21877 T21878 det a,strategy
R6488 T21878 T21868 nsubjpass strategy,developed
R6489 T21879 T21878 amod nested,strategy
R6490 T21880 T21878 compound PCR,strategy
R6491 T21881 T21868 auxpass was,developed
R6492 T21882 T21868 prep in,developed
R6493 T21883 T21882 pobj order,in
R6494 T21884 T21885 aux to,genotype
R6495 T21885 T21883 acl genotype,order
R6496 T21886 T21887 amod pre-implantation,embryos
R6497 T21887 T21885 dobj embryos,genotype
R6498 T21888 T21868 punct .,developed
R6499 T21890 T21891 amod Separate,reactions
R6500 T21891 T21892 nsubjpass reactions,used
R6501 T21893 T21892 auxpass were,used
R6502 T21894 T21895 aux to,amplify
R6503 T21895 T21892 advcl amplify,used
R6504 T21896 T21897 det a,fragment
R6505 T21897 T21895 dobj fragment,amplify
R6506 T21898 T21899 nummod 429,bp
R6507 T21899 T21897 compound bp,fragment
R6508 T21900 T21897 prep from,fragment
R6509 T21901 T21902 det the,allele
R6510 T21902 T21900 pobj allele,from
R6511 T21903 T21904 amod wild,type
R6512 T21904 T21902 compound type,allele
R6513 T21905 T21904 punct -,type
R6514 T21906 T21897 cc and,fragment
R6515 T21907 T21908 det a,segment
R6516 T21908 T21897 conj segment,fragment
R6517 T21909 T21910 nummod 389,bp
R6518 T21910 T21908 compound bp,segment
R6519 T21911 T21908 prep from,segment
R6520 T21912 T21913 det the,allele
R6521 T21913 T21911 pobj allele,from
R6522 T21914 T21913 compound mutant,allele
R6523 T21915 T21897 punct ", ",fragment
R6524 T21916 T21897 appos both,fragment
R6525 T21917 T21916 acl spanning,both
R6526 T21918 T21919 det the,end
R6527 T21919 T21917 dobj end,spanning
R6528 T21920 T21919 nummod 3,end
R6529 T21921 T21920 punct ',3
R6530 T21922 T21919 prep of,end
R6531 T21923 T21924 det the,arm
R6532 T21924 T21922 pobj arm,of
R6533 T21925 T21924 amod short,arm
R6534 T21926 T21927 punct (,upstream
R6535 T21927 T21924 parataxis upstream,arm
R6536 T21928 T21927 punct ),upstream
R6537 T21929 T21924 prep of,arm
R6538 T21930 T21931 det the,vector
R6539 T21931 T21929 pobj vector,of
R6540 T21932 T21931 compound targeting,vector
R6541 T21933 T21892 punct .,used
R6542 T21935 T21936 prep In,co-amplified
R6543 T21937 T21938 det both,reactions
R6544 T21938 T21935 pobj reactions,In
R6545 T21939 T21936 punct ", ",co-amplified
R6546 T21940 T21941 det a,sequence
R6547 T21941 T21936 nsubjpass sequence,co-amplified
R6548 T21942 T21943 nummod 213,bp
R6549 T21943 T21941 compound bp,sequence
R6550 T21944 T21941 acl located,sequence
R6551 T21945 T21944 prep within,located
R6552 T21946 T21947 det the,arm
R6553 T21947 T21945 pobj arm,within
R6554 T21948 T21947 amod short,arm
R6555 T21949 T21936 auxpass was,co-amplified
R6556 T21950 T21936 prep with,co-amplified
R6557 T21951 T21952 det the,products
R6558 T21952 T21950 pobj products,with
R6559 T21953 T21952 punct ',products
R6560 T21954 T21952 amod diagnostic,products
R6561 T21955 T21952 punct ',products
R6562 T21956 T21936 prep as,co-amplified
R6563 T21957 T21958 det an,control
R6564 T21958 T21956 pobj control,as
R6565 T21959 T21958 amod internal,control
R6566 T21960 T21936 punct .,co-amplified
R6567 T21962 T21963 det The,primers
R6568 T21963 T21966 nsubjpass primers,used
R6569 T21964 T21963 amod same,primers
R6570 T21965 T21963 compound sense,primers
R6571 T21967 T21966 auxpass were,used
R6572 T21968 T21969 aux to,amplify
R6573 T21969 T21966 advcl amplify,used
R6574 T21970 T21969 punct ',amplify
R6575 T21971 T21972 amod diagnostic,sequences
R6576 T21972 T21969 dobj sequences,amplify
R6577 T21973 T21972 punct ',sequences
R6578 T21974 T21972 prep in,sequences
R6579 T21975 T21976 det both,reactions
R6580 T21976 T21974 pobj reactions,in
R6581 T21977 T21969 punct ", ",amplify
R6582 T21978 T21979 mark whereas,were
R6583 T21979 T21969 advcl were,amplify
R6584 T21980 T21981 det the,primers
R6585 T21981 T21979 nsubj primers,were
R6586 T21982 T21981 compound antisense,primers
R6587 T21983 T21984 npadvmod allele,specific
R6588 T21984 T21979 acomp specific,were
R6589 T21985 T21984 punct -,specific
R6590 T21986 T21966 punct .,used
R6591 T21988 T21989 det The,primers
R6592 T21989 T21990 nsubjpass primers,depicted
R6593 T21991 T21989 punct ", ",primers
R6594 T21992 T21989 acl represented,primers
R6595 T21993 T21992 agent by,represented
R6596 T21994 T21993 pobj triangles,by
R6597 T21995 T21990 punct ", ",depicted
R6598 T21996 T21990 auxpass are,depicted
R6599 T21997 T21990 prep in,depicted
R6600 T21998 T21999 nummod two,sets
R6601 T21999 T21997 pobj sets,in
R6602 T22000 T21999 punct (,sets
R6603 T22001 T21999 amod nested,sets
R6604 T22002 T21999 punct ),sets
R6605 T22003 T21999 prep for,sets
R6606 T22004 T22003 pobj each,for
R6607 T22005 T22004 prep of,each
R6608 T22006 T22007 det the,reactions
R6609 T22007 T22005 pobj reactions,of
R6610 T22008 T22007 nummod three,reactions
R6611 T22009 T21990 punct .,depicted
R6612 T22011 T22012 nsubjpass Exons,represented
R6613 T22012 T22014 ccomp represented,cassette
R6614 T22013 T22012 auxpass are,represented
R6615 T22015 T22012 agent by,represented
R6616 T22016 T22017 amod open,rectangles
R6617 T22017 T22015 pobj rectangles,by
R6618 T22018 T22017 punct ", ",rectangles
R6619 T22019 T22017 amod vertical,rectangles
R6620 T22020 T22014 punct ", ",cassette
R6621 T22021 T22014 det the,cassette
R6622 T22022 T22023 compound PGK,Neo
R6623 T22023 T22014 compound Neo,cassette
R6624 T22024 T22023 punct -,Neo
R6625 T22025 T22014 agent by,cassette
R6626 T22026 T22027 det an,rectangle
R6627 T22027 T22025 pobj rectangle,by
R6628 T22028 T22027 amod open,rectangle
R6629 T22029 T22027 punct ", ",rectangle
R6630 T22030 T22027 amod horizontal,rectangle
R6631 T22031 T22014 punct ", ",cassette
R6632 T22032 T22033 mark while,denote
R6633 T22033 T22014 advcl denote,cassette
R6634 T22034 T22035 amod thin,lines
R6635 T22035 T22033 nsubj lines,denote
R6636 T22036 T22035 amod vertical,lines
R6637 T22037 T22038 det the,boundaries
R6638 T22038 T22033 dobj boundaries,denote
R6639 T22039 T22038 prep of,boundaries
R6640 T22040 T22041 det the,arm
R6641 T22041 T22039 pobj arm,of
R6642 T22042 T22041 amod short,arm
R6643 T22043 T22038 cc and,boundaries
R6644 T22044 T22045 det the,end
R6645 T22045 T22038 conj end,boundaries
R6646 T22046 T22045 nummod 5,end
R6647 T22047 T22046 punct ',5
R6648 T22048 T22045 prep of,end
R6649 T22049 T22050 det the,arm
R6650 T22050 T22048 pobj arm,of
R6651 T22051 T22050 amod long,arm
R6652 T22052 T22050 punct (,arm
R6653 T22053 T22050 amod downstream,arm
R6654 T22054 T22050 punct ),arm
R6655 T22055 T22014 punct .,cassette
R6656 T22057 T22058 det A,rectangle
R6657 T22058 T22062 nsubj rectangle,delineates
R6658 T22059 T22058 amod grey,rectangle
R6659 T22060 T22058 punct ", ",rectangle
R6660 T22061 T22058 amod horizontal,rectangle
R6661 T22063 T22064 det the,segment
R6662 T22064 T22062 dobj segment,delineates
R6663 T22065 T22064 prep of,segment
R6664 T22066 T22067 det the,allele
R6665 T22067 T22065 pobj allele,of
R6666 T22068 T22069 amod wild,type
R6667 T22069 T22067 compound type,allele
R6668 T22070 T22069 punct -,type
R6669 T22071 T22072 dep that,replaced
R6670 T22072 T22064 relcl replaced,segment
R6671 T22073 T22072 auxpass is,replaced
R6672 T22074 T22072 agent by,replaced
R6673 T22075 T22076 det the,cassette
R6674 T22076 T22074 pobj cassette,by
R6675 T22077 T22078 compound PGK,Neo
R6676 T22078 T22076 compound Neo,cassette
R6677 T22079 T22078 punct -,Neo
R6678 T22080 T22072 prep in,replaced
R6679 T22081 T22082 det the,allele
R6680 T22082 T22080 pobj allele,in
R6681 T22083 T22082 compound mutant,allele
R6682 T22084 T22062 punct .,delineates
R6683 T22641 T22640 prep of,Genotyping
R6684 T22642 T22643 amod pre-implantation,embryos
R6685 T22643 T22641 pobj embryos,of
R6686 T22644 T22640 punct .,Genotyping
R6687 T22646 T22647 det A,strategy
R6688 T22647 T22650 nsubjpass strategy,used
R6689 T22648 T22647 amod nested,strategy
R6690 T22649 T22647 compound PCR,strategy
R6691 T22651 T22650 auxpass was,used
R6692 T22652 T22653 aux to,genotype
R6693 T22653 T22650 advcl genotype,used
R6694 T22654 T22653 dobj embryos,genotype
R6695 T22655 T22656 amod prior,to
R6696 T22656 T22653 prep to,genotype
R6697 T22657 T22656 pobj implantation,to
R6698 T22658 T22659 mark as,described
R6699 T22659 T22653 advcl described,genotype
R6700 T22660 T22659 prep in,described
R6701 T22661 T22660 pobj Figure,in
R6702 T22662 T22661 nummod 4,Figure
R6703 T22663 T22650 punct .,used
R6704 T22665 T22666 nmod Capn2,mice
R6705 T22666 T22670 nsubjpass mice,mated
R6706 T22667 T22665 punct +,Capn2
R6707 T22668 T22665 punct /,Capn2
R6708 T22669 T22665 punct -,Capn2
R6709 T22671 T22670 auxpass were,mated
R6710 T22672 T22670 cc and,mated
R6711 T22673 T22674 det the,date
R6712 T22674 T22675 nsubj date,established
R6713 T22675 T22670 conj established,mated
R6714 T22676 T22674 prep of,date
R6715 T22677 T22676 pobj fertilization,of
R6716 T22678 T22675 agent by,established
R6717 T22679 T22680 det the,appearance
R6718 T22680 T22678 pobj appearance,by
R6719 T22681 T22680 prep of,appearance
R6720 T22682 T22683 amod vaginal,plugs
R6721 T22683 T22681 pobj plugs,of
R6722 T22684 T22675 punct .,established
R6723 T22686 T22687 nmod Blastocyst,embryos
R6724 T22687 T22695 nsubjpass embryos,flushed
R6725 T22688 T22689 punct (,E3.5
R6726 T22689 T22686 parataxis E3.5,Blastocyst
R6727 T22690 T22689 punct ),E3.5
R6728 T22691 T22686 cc or,Blastocyst
R6729 T22692 T22693 nummod 8,cell
R6730 T22693 T22686 conj cell,Blastocyst
R6731 T22694 T22693 punct -,cell
R6732 T22696 T22697 punct (,E2.5
R6733 T22697 T22687 parataxis E2.5,embryos
R6734 T22698 T22697 punct ),E2.5
R6735 T22699 T22695 auxpass were,flushed
R6736 T22700 T22695 prep from,flushed
R6737 T22701 T22702 det the,oviducts
R6738 T22702 T22700 pobj oviducts,from
R6739 T22703 T22695 cc and,flushed
R6740 T22704 T22705 advmod then,digested
R6741 T22705 T22695 conj digested,flushed
R6742 T22706 T22705 prep with,digested
R6743 T22707 T22708 compound proteinase,K
R6744 T22708 T22706 pobj K,with
R6745 T22709 T22695 punct .,flushed
R6746 T22711 T22712 prep In,co-amplified
R6747 T22713 T22714 amod separate,segments
R6748 T22714 T22711 pobj segments,In
R6749 T22715 T22714 compound reactions,segments
R6750 T22716 T22714 acl found,segments
R6751 T22717 T22716 advmod exclusively,found
R6752 T22718 T22716 prep in,found
R6753 T22719 T22720 preconj either,type
R6754 T22720 T22718 pobj type,in
R6755 T22721 T22720 det the,type
R6756 T22722 T22720 amod wild,type
R6757 T22723 T22720 punct -,type
R6758 T22724 T22720 cc or,type
R6759 T22725 T22720 conj mutant,type
R6760 T22726 T22712 nsubjpass alleles,co-amplified
R6761 T22727 T22712 auxpass were,co-amplified
R6762 T22728 T22712 prep with,co-amplified
R6763 T22729 T22730 det an,sequence
R6764 T22730 T22728 pobj sequence,with
R6765 T22731 T22730 amod internal,sequence
R6766 T22732 T22730 compound control,sequence
R6767 T22733 T22730 punct ", ",sequence
R6768 T22734 T22730 acl located,sequence
R6769 T22735 T22734 prep in,located
R6770 T22736 T22737 det the,arm
R6771 T22737 T22735 pobj arm,in
R6772 T22738 T22737 amod short,arm
R6773 T22739 T22737 punct (,arm
R6774 T22740 T22737 amod upstream,arm
R6775 T22741 T22737 punct ),arm
R6776 T22742 T22737 prep of,arm
R6777 T22743 T22744 det the,vector
R6778 T22744 T22742 pobj vector,of
R6779 T22745 T22744 compound targeting,vector
R6780 T22746 T22744 punct ", ",vector
R6781 T22747 T22748 dep which,found
R6782 T22748 T22744 relcl found,vector
R6783 T22749 T22748 auxpass is,found
R6784 T22750 T22748 prep in,found
R6785 T22751 T22752 det both,alleles
R6786 T22752 T22750 pobj alleles,in
R6787 T22753 T22712 punct .,co-amplified
R6788 T22755 T22756 det The,products
R6789 T22756 T22758 nsubj products,were
R6790 T22757 T22756 amod final,products
R6791 T22759 T22760 nummod 429,bp
R6792 T22760 T22758 attr bp,were
R6793 T22761 T22758 prep for,were
R6794 T22762 T22763 det the,allele
R6795 T22763 T22761 pobj allele,for
R6796 T22764 T22765 amod wild,type
R6797 T22765 T22763 compound type,allele
R6798 T22766 T22765 punct -,type
R6799 T22767 T22758 punct ", ",were
R6800 T22768 T22769 nummod 389,bp
R6801 T22769 T22758 conj bp,were
R6802 T22770 T22769 prep for,bp
R6803 T22771 T22772 det the,allele
R6804 T22772 T22770 pobj allele,for
R6805 T22773 T22772 compound mutant,allele
R6806 T22774 T22769 punct ", ",bp
R6807 T22775 T22769 cc and,bp
R6808 T22776 T22777 nummod 213,bp
R6809 T22777 T22769 conj bp,bp
R6810 T22778 T22777 prep for,bp
R6811 T22779 T22780 det the,control
R6812 T22780 T22778 pobj control,for
R6813 T22781 T22780 amod internal,control
R6814 T22782 T22758 punct .,were
R6815 T22784 T22785 punct (,A
R6816 T22785 T22786 meta A,example
R6817 T22787 T22785 punct ),A
R6818 T22788 T22786 det A,example
R6819 T22789 T22786 amod representative,example
R6820 T22790 T22786 prep of,example
R6821 T22791 T22792 det the,genotyping
R6822 T22792 T22790 pobj genotyping,of
R6823 T22793 T22792 prep of,genotyping
R6824 T22794 T22795 compound blastocyst,embryos
R6825 T22795 T22793 pobj embryos,of
R6826 T22796 T22795 compound stage,embryos
R6827 T22797 T22786 punct .,example
R6828 T22799 T22800 nmod Embryos,1
R6829 T22800 T22802 nsubj 1,were
R6830 T22801 T22800 punct #,1
R6831 T22803 T22800 punct ", ",1
R6832 T22804 T22800 conj 2,1
R6833 T22805 T22804 punct ", ",2
R6834 T22806 T22804 conj 4,2
R6835 T22807 T22806 punct ", ",4
R6836 T22808 T22806 conj 5,4
R6837 T22809 T22808 punct ", ",5
R6838 T22810 T22808 cc and,5
R6839 T22811 T22808 conj 6,5
R6840 T22812 T22802 attr Capn2,were
R6841 T22813 T22812 punct +,Capn2
R6842 T22814 T22812 punct /,Capn2
R6843 T22815 T22812 punct -,Capn2
R6844 T22816 T22817 mark whereas,were
R6845 T22817 T22802 advcl were,were
R6846 T22818 T22819 nmod embryos,3
R6847 T22819 T22817 nsubj 3,were
R6848 T22820 T22819 punct #,3
R6849 T22821 T22819 cc and,3
R6850 T22822 T22823 punct #,7
R6851 T22823 T22819 conj 7,3
R6852 T22824 T22817 attr Capn2,were
R6853 T22825 T22824 punct +,Capn2
R6854 T22826 T22824 punct /,Capn2
R6855 T22827 T22824 punct +,Capn2
R6856 T22828 T22817 punct ", ",were
R6857 T22829 T22817 advcl denoted,were
R6858 T22830 T22829 agent by,denoted
R6859 T22831 T22832 det the,absence
R6860 T22832 T22830 pobj absence,by
R6861 T22833 T22832 prep of,absence
R6862 T22834 T22835 det the,signal
R6863 T22835 T22833 pobj signal,of
R6864 T22836 T22837 nummod 389,bp
R6865 T22837 T22835 compound bp,signal
R6866 T22838 T22835 compound mutant,signal
R6867 T22839 T22802 punct .,were
R6868 T22841 T22842 punct (,B
R6869 T22842 T22843 meta B,shown
R6870 T22844 T22842 punct ),B
R6871 T22845 T22846 det An,example
R6872 T22846 T22843 nsubjpass example,shown
R6873 T22847 T22846 prep of,example
R6874 T22848 T22849 det the,genotyping
R6875 T22849 T22847 pobj genotyping,of
R6876 T22850 T22849 prep of,genotyping
R6877 T22851 T22852 nummod 8,cell
R6878 T22852 T22854 compound cell,embryos
R6879 T22853 T22852 punct -,cell
R6880 T22854 T22850 pobj embryos,of
R6881 T22855 T22843 auxpass is,shown
R6882 T22856 T22843 punct .,shown
R6883 T22858 T22859 nmod Embryos,1
R6884 T22859 T22861 nsubj 1,were
R6885 T22860 T22859 punct #,1
R6886 T22862 T22859 punct ", ",1
R6887 T22863 T22859 conj 2,1
R6888 T22864 T22863 punct ", ",2
R6889 T22865 T22863 conj 3,2
R6890 T22866 T22865 punct ", ",3
R6891 T22867 T22865 conj 5,3
R6892 T22868 T22867 punct ", ",5
R6893 T22869 T22867 cc and,5
R6894 T22870 T22867 conj 6,5
R6895 T22871 T22861 attr Capn2,were
R6896 T22872 T22871 punct +,Capn2
R6897 T22873 T22871 punct /,Capn2
R6898 T22874 T22871 punct -,Capn2
R6899 T22875 T22876 mark while,was
R6900 T22876 T22861 advcl was,were
R6901 T22877 T22876 nsubj embryo,was
R6902 T22878 T22877 punct #,embryo
R6903 T22879 T22877 nummod 4,embryo
R6904 T22880 T22876 attr Capn2,was
R6905 T22881 T22880 punct -,Capn2
R6906 T22882 T22880 punct /,Capn2
R6907 T22883 T22880 punct -,Capn2
R6908 T22884 T22880 punct ", ",Capn2
R6909 T22885 T22880 acl marked,Capn2
R6910 T22886 T22885 agent by,marked
R6911 T22887 T22888 det the,absence
R6912 T22888 T22886 pobj absence,by
R6913 T22889 T22888 prep of,absence
R6914 T22890 T22891 det the,signal
R6915 T22891 T22889 pobj signal,of
R6916 T22892 T22893 nummod 429,bp
R6917 T22893 T22891 nmod bp,signal
R6918 T22894 T22895 amod wild,type
R6919 T22895 T22891 compound type,signal
R6920 T22896 T22895 punct -,type
R6921 T22897 T22861 punct .,were
R6922 T22899 T22900 punct (,M
R6923 T22900 T22901 nsubj M,denotes
R6924 T22902 T22900 punct ),M
R6925 T22903 T22904 det the,marker
R6926 T22904 T22901 dobj marker,denotes
R6927 T22905 T22906 amod molecular,weight
R6928 T22906 T22904 compound weight,marker
R6929 T22907 T22901 punct .,denotes
R6930 T22936 T22937 compound Genotype,distribution
R6931 T22938 T22937 prep of,distribution
R6932 T22939 T22938 pobj offspring,of
R6933 T22940 T22939 acl derived,offspring
R6934 T22941 T22940 prep from,derived
R6935 T22942 T22943 nmod Capn2,mice
R6936 T22943 T22941 pobj mice,from
R6937 T22944 T22943 amod transgenic,mice
R6938 T22945 T22937 punct .,distribution
R1881 T7005 T7006 det "Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The",genotyping
R1882 T7006 T7007 nsubjpass genotyping,shown
R1883 T7008 T7006 nmod results,genotyping
R1884 T7009 T7006 prep for,genotyping
R1885 T7010 T7009 pobj weanlings,for
R1886 T7011 T7010 punct ", ",weanlings
R1887 T7012 T7013 nmod post-implantation,embryos
R1888 T7013 T7010 appos embryos,weanlings
R1889 T7014 T7012 punct ", ",post-implantation
R1890 T7015 T7012 cc and,post-implantation
R1891 T7016 T7012 conj pre-implantation,post-implantation

craft-ca-core-dev

Below, discontinuous spans are shown in the chain model. You can change it to the bag model.

Id Subject Object Predicate Lexical cue
T22582 2803-2805 SO:0000028 denotes bp
T22581 2760-2765 PR:000005015 denotes Capn2
T22580 2746-2752 UBERON:0000922 denotes embryo
T22579 2731-2736 PR:000005015 denotes Capn2
T22578 2699-2706 UBERON:0000922 denotes Embryos
T22577 2674-2688 UBERON:0019252 denotes 8-cell embryos
T22576 2620-2622 SO:0000028 denotes bp
T22575 2576-2581 PR:000005015 denotes Capn2
T22574 2553-2560 UBERON:0000922 denotes embryos
T22573 2536-2541 PR:000005015 denotes Capn2
T22572 2504-2511 UBERON:0000922 denotes Embryos
T22571 2495-2502 UBERON:0000922 denotes embryos
T22570 2478-2488 UBERON:0000358 denotes blastocyst
T22569 2399-2401 SO:0000028 denotes bp
T22568 2383-2389 SO:0001023 denotes allele
T22567 2365-2367 SO:0000028 denotes bp
T22566 2353-2359 SO:0001023 denotes allele
T22565 2332-2334 SO:0000028 denotes bp
T22564 2295-2302 SO:0001023 denotes alleles
T22563 2254-2270 SO:0001644 denotes targeting vector
T22562 2150-2157 SO:0001023 denotes alleles
T22561 2021-2029 UBERON:0000993 denotes oviducts
T22560 1977-1991 UBERON:0019252 denotes 8-cell embryos
T22559 1956-1966 UBERON:0000358 denotes Blastocyst
T22558 1941-1954 UBERON:0010148 denotes vaginal plugs
T22557 1894-1907 GO:0009566 denotes fertilization
T22556 1872-1877 GO:0007618 denotes mated
T22555 1862-1866 NCBITaxon:10088 denotes mice
T22554 1853-1858 PR:000005015 denotes Capn2
T22553 1814-1826 GO:0007566 denotes implantation
T22552 1797-1804 UBERON:0000922 denotes embryos
T22551 1745-1752 UBERON:0000922 denotes embryos
T22550 1732-1744 GO:0007566 denotes implantation
T21822 1696-1702 SO:0001023 denotes allele
T21821 1673-1681 SO:0005853 denotes cassette
T21820 1634-1640 SO:0001023 denotes allele
T21819 1412-1420 SO:0005853 denotes cassette
T21818 1348-1353 SO:0000147 denotes Exons
T21817 1246-1253 SO:0000112 denotes primers
T21816 1225-1231 SO:0001023 denotes allele
T21815 1212-1219 SO:0000112 denotes primers
T21814 1202-1211 SO:0000077 denotes antisense
T21813 1119-1126 SO:0000112 denotes primers
T21812 991-993 SO:0000028 denotes bp
T21811 948-964 SO:0001644 denotes targeting vector
T21810 880-886 SO:0001023 denotes allele
T21809 853-855 SO:0000028 denotes bp
T21808 836-842 SO:0001023 denotes allele
T21807 805-807 SO:0000028 denotes bp
T21806 750-757 UBERON:0000922 denotes embryos
T21805 737-749 GO:0007566 denotes implantation
T21804 658-665 SO:0000704 denotes genetic
T21803 614-621 UBERON:0000922 denotes embryos
T21802 601-613 GO:0007566 denotes implantation
T22930 58-63 PR:000005015 denotes Capn2
T22931 75-79 NCBITaxon:10088 denotes mice
T6435 2943-2950 UBERON:0000922 denotes embryos
T6434 2930-2942 GO:0007566 denotes implantation
T6433 2908-2920 GO:0007566 denotes implantation

craft-ca-core-ex-dev

Below, discontinuous spans are shown in the chain model. You can change it to the bag model.

Id Subject Object Predicate Lexical cue
T22639 2857-2863 CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag denotes marker
T22638 2840-2849 CHEBI_EXT:polyatomic_entity_or_group denotes molecular
T22637 2825-2826 CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag denotes M
T22636 2806-2815 SO_EXT:wild_type_entity_or_quality denotes wild-type
T22635 2803-2805 SO_EXT:0000028 denotes bp
T22634 2767-2768 SO_EXT:sequence_nullness_or_absence denotes -
T22633 2765-2766 SO_EXT:sequence_nullness_or_absence denotes -
T22632 2760-2765 PR_EXT:000005015 denotes Capn2
T22631 2746-2752 UBERON:0000922 denotes embryo
T22630 2738-2739 SO_EXT:sequence_nullness_or_absence denotes -
T22629 2736-2737 SO_EXT:normal_or_wild_type_or_present denotes +
T22628 2731-2736 PR_EXT:000005015 denotes Capn2
T22627 2699-2706 UBERON:0000922 denotes Embryos
T22626 2676-2680 CL_GO_EXT:cell denotes cell
T22625 2674-2688 UBERON:0019252 denotes 8-cell embryos
T22624 2623-2629 SO_EXT:sequence_altered_entity_or_alteration_process denotes mutant
T22623 2620-2622 SO_EXT:0000028 denotes bp
T22622 2583-2584 SO_EXT:normal_or_wild_type_or_present denotes +
T22621 2581-2582 SO_EXT:normal_or_wild_type_or_present denotes +
T22620 2576-2581 PR_EXT:000005015 denotes Capn2
T22603 2233-2241 SO_EXT:sequence_upstreamness denotes upstream
T22602 2201-2209 SO_EXT:biological_sequence denotes sequence
T22601 2150-2157 SO_EXT:0001023 denotes alleles
T22600 2143-2149 SO_EXT:sequence_altered_entity_or_alteration_process denotes mutant
T22599 2130-2139 SO_EXT:wild_type_entity_or_quality denotes wild-type
T22598 2053-2063 GO_EXT:0008233 denotes proteinase
T22597 2021-2029 UBERON:0000993 denotes oviducts
T22596 1979-1983 CL_GO_EXT:cell denotes cell
T22595 1977-1991 UBERON:0019252 denotes 8-cell embryos
T22594 1956-1966 UBERON:0000358 denotes Blastocyst
T22593 1941-1954 UBERON:0010148 denotes vaginal plugs
T22592 1894-1907 GO:0009566 denotes fertilization
T22591 1872-1877 GO:0007618 denotes mated
T22590 1862-1866 NCBITaxon:10088 denotes mice
T22589 1860-1861 SO_EXT:sequence_nullness_or_absence denotes -
T22588 1858-1859 SO_EXT:normal_or_wild_type_or_present denotes +
T22587 1853-1858 PR_EXT:000005015 denotes Capn2
T22586 1814-1826 GO:0007566 denotes implantation
T22585 1797-1804 UBERON:0000922 denotes embryos
T22584 1745-1752 UBERON:0000922 denotes embryos
T22583 1732-1744 GO:0007566 denotes implantation
T21834 927-935 SO_EXT:sequence_upstreamness denotes upstream
T21833 880-886 SO_EXT:0001023 denotes allele
T21832 873-879 SO_EXT:sequence_altered_entity_or_alteration_process denotes mutant
T21831 853-855 SO_EXT:0000028 denotes bp
T21830 836-842 SO_EXT:0001023 denotes allele
T21829 826-835 SO_EXT:wild_type_entity_or_quality denotes wild-type
T21828 805-807 SO_EXT:0000028 denotes bp
T21827 750-757 UBERON:0000922 denotes embryos
T21826 737-749 GO:0007566 denotes implantation
T21825 658-665 SO_EXT:0000704 denotes genetic
T21824 614-621 UBERON:0000922 denotes embryos
T21823 601-613 GO:0007566 denotes implantation
T21856 1696-1702 SO_EXT:0001023 denotes allele
T21855 1689-1695 SO_EXT:sequence_altered_entity_or_alteration_process denotes mutant
T21854 1673-1681 SO_EXT:0005853 denotes cassette
T21853 1669-1672 CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase denotes Neo
T21852 1665-1668 GO_EXT:phosphoglycerate_kinase denotes PGK
T21851 1649-1657 SO_EXT:sequence_substitution_process denotes replaced
T21850 1634-1640 SO_EXT:0001023 denotes allele
T21849 1624-1633 SO_EXT:wild_type_entity_or_quality denotes wild-type
T21848 1548-1558 SO_EXT:sequence_downstreamness denotes downstream
T21847 1412-1420 SO_EXT:0005853 denotes cassette
T21846 1408-1411 CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase denotes Neo
T21845 1404-1407 GO_EXT:phosphoglycerate_kinase denotes PGK
T21844 1348-1353 SO_EXT:0000147 denotes Exons
T21843 1246-1253 SO_EXT:0000112 denotes primers
T21842 1225-1231 SO_EXT:0001023 denotes allele
T21841 1212-1219 SO_EXT:0000112 denotes primers
T21840 1202-1211 SO:0000077 denotes antisense
T21839 1161-1170 SO_EXT:biological_sequence denotes sequences
T21838 1119-1126 SO_EXT:0000112 denotes primers
T21837 994-1002 SO_EXT:biological_sequence denotes sequence
T21836 991-993 SO_EXT:0000028 denotes bp
T21835 948-964 SO_EXT:0001644 denotes targeting vector
T22619 2553-2560 UBERON:0000922 denotes embryos
T22618 2543-2544 SO_EXT:sequence_nullness_or_absence denotes -
T22617 2541-2542 SO_EXT:normal_or_wild_type_or_present denotes +
T22616 2536-2541 PR_EXT:000005015 denotes Capn2
T22615 2504-2511 UBERON:0000922 denotes Embryos
T22614 2495-2502 UBERON:0000922 denotes embryos
T22613 2478-2488 UBERON:0000358 denotes blastocyst
T22612 2399-2401 SO_EXT:0000028 denotes bp
T22611 2383-2389 SO_EXT:0001023 denotes allele
T22610 2376-2382 SO_EXT:sequence_altered_entity_or_alteration_process denotes mutant
T22609 2365-2367 SO_EXT:0000028 denotes bp
T22608 2353-2359 SO_EXT:0001023 denotes allele
T22607 2343-2352 SO_EXT:wild_type_entity_or_quality denotes wild-type
T22606 2332-2334 SO_EXT:0000028 denotes bp
T22605 2295-2302 SO_EXT:0001023 denotes alleles
T22604 2254-2270 SO_EXT:0001644 denotes targeting vector
T22932 10-18 SO_EXT:genotype_or_entity_with_genotype denotes Genotype
T22933 58-63 PR_EXT:000005015 denotes Capn2
T22934 64-74 SO_EXT:transgenic_entity denotes transgenic
T22935 75-79 NCBITaxon:10088 denotes mice
T6558 2943-2950 UBERON:0000922 denotes embryos
T6557 2930-2942 GO:0007566 denotes implantation
T6556 2908-2920 GO:0007566 denotes implantation