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PMC:1482699 / 43350-45368 JSONTXT 4 Projects

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Id Subject Object Predicate Lexical cue
T26350 0-9 NN denotes Candidate
T26351 10-14 NN denotes gene
T26352 15-25 NN denotes sequencing
T26353 25-264 sentence denotes PCR amplicons covering the coding sequence and partial 5' and 3' untranslated regions for each selected candidate gene were amplified, purified and sequenced from the CAST strain using protocols outlined in [57] with slight modifications.
T26354 26-29 NN denotes PCR
T26355 30-39 NNS denotes amplicons
T26357 40-48 VBG denotes covering
T26358 49-52 DT denotes the
T26360 53-59 NN denotes coding
T26359 60-68 NN denotes sequence
T26361 69-72 CC denotes and
T26362 73-80 JJ denotes partial
T26364 81-82 CD denotes 5
T26365 82-83 SYM denotes '
T26366 84-87 CC denotes and
T26367 88-89 CD denotes 3
T26368 89-90 SYM denotes '
T26369 91-103 JJ denotes untranslated
T26363 104-111 NNS denotes regions
T26370 112-115 IN denotes for
T26371 116-120 DT denotes each
T26373 121-129 VBN denotes selected
T26374 130-139 NN denotes candidate
T26372 140-144 NN denotes gene
T26375 145-149 VBD denotes were
T26356 150-159 VBN denotes amplified
T26376 159-161 , denotes ,
T26377 161-169 VBN denotes purified
T26378 170-173 CC denotes and
T26379 174-183 VBN denotes sequenced
T26380 184-188 IN denotes from
T26381 189-192 DT denotes the
T26383 193-197 NN denotes CAST
T26382 198-204 NN denotes strain
T26384 205-210 VBG denotes using
T26385 211-220 NNS denotes protocols
T26386 221-229 VBN denotes outlined
T26387 230-232 IN denotes in
T26388 233-234 -LRB- denotes [
T26389 234-236 CD denotes 57
T26390 236-237 -RRB- denotes ]
T26391 238-242 IN denotes with
T26392 243-249 JJ denotes slight
T26393 250-263 NNS denotes modifications
T26394 263-264 . denotes .
T26395 264-383 sentence denotes Total RNA from brain, liver, spleen, lung and testis was isolated from an adult CAST male mouse using Trizol (Ambion).
T26396 265-270 JJ denotes Total
T26397 271-274 NN denotes RNA
T26399 275-279 IN denotes from
T26400 280-285 NN denotes brain
T26401 285-287 , denotes ,
T26402 287-292 NN denotes liver
T26403 292-294 , denotes ,
T26404 294-300 NN denotes spleen
T26405 300-302 , denotes ,
T26406 302-306 NN denotes lung
T26407 307-310 CC denotes and
T26408 311-317 NN denotes testis
T26409 318-321 VBD denotes was
T26398 322-330 VBN denotes isolated
T26410 331-335 IN denotes from
T26411 336-338 DT denotes an
T26413 339-344 JJ denotes adult
T26414 345-349 NN denotes CAST
T26415 350-354 JJ denotes male
T26412 355-360 NN denotes mouse
T26416 361-366 VBG denotes using
T26417 367-373 NN denotes Trizol
T26418 374-375 -LRB- denotes (
T26419 375-381 NNP denotes Ambion
T26420 381-382 -RRB- denotes )
T26421 382-383 . denotes .
T26422 383-443 sentence denotes cDNA was produced from total RNA using standard procedures.
T26423 384-388 NN denotes cDNA
T26425 389-392 VBD denotes was
T26424 393-401 VBN denotes produced
T26426 402-406 IN denotes from
T26427 407-412 JJ denotes total
T26428 413-416 NN denotes RNA
T26429 417-422 VBG denotes using
T26430 423-431 JJ denotes standard
T26431 432-442 NNS denotes procedures
T26432 442-443 . denotes .
T26433 443-527 sentence denotes PCR primer sets for each gene were designed using Primer3 [58] (Additional File 5).
T26434 444-447 NN denotes PCR
T26436 448-454 NN denotes primer
T26435 455-459 NNS denotes sets
T26438 460-463 IN denotes for
T26439 464-468 DT denotes each
T26440 469-473 NN denotes gene
T26441 474-478 VBD denotes were
T26437 479-487 VBN denotes designed
T26442 488-493 VBG denotes using
T26443 494-501 NN denotes Primer3
T26444 502-503 -LRB- denotes [
T26445 503-505 CD denotes 58
T26446 505-506 -RRB- denotes ]
T26447 507-508 -LRB- denotes (
T26449 508-518 JJ denotes Additional
T26448 519-523 NN denotes File
T26450 524-525 CD denotes 5
T26451 525-526 -RRB- denotes )
T26452 526-527 . denotes .
T26453 527-663 sentence denotes The initial amplification was performed in 10 μl PCR reactions using the Invitrogen Platinum TaqPCRx amplification system (Invitrogen).
T26454 528-531 DT denotes The
T26456 532-539 JJ denotes initial
T26455 540-553 NN denotes amplification
T26458 554-557 VBD denotes was
T26457 558-567 VBN denotes performed
T26459 568-570 IN denotes in
T26460 571-573 CD denotes 10
T26461 574-576 NN denotes μl
T26463 577-580 NN denotes PCR
T26462 581-590 NNS denotes reactions
T26464 591-596 VBG denotes using
T26465 597-600 DT denotes the
T26467 601-611 NNP denotes Invitrogen
T26468 612-620 NN denotes Platinum
T26469 621-628 NN denotes TaqPCRx
T26470 629-642 NN denotes amplification
T26466 643-649 NN denotes system
T26471 650-651 -LRB- denotes (
T26472 651-661 NNP denotes Invitrogen
T26473 661-662 -RRB- denotes )
T26474 662-663 . denotes .
T26475 663-842 sentence denotes The reactions contained 1X PCR buffer, 1X Enhancer solution, 1.5 mM MgSO4, 0.17 mM dNTPs, 1 μM each primer, 0.1 unit of Platinum Taq (Invitrogen) and approximately 25 ng of cDNA.
T26476 664-667 DT denotes The
T26477 668-677 NNS denotes reactions
T26478 678-687 VBD denotes contained
T26479 688-690 NN denotes 1X
T26481 691-694 NN denotes PCR
T26480 695-701 NN denotes buffer
T26482 701-703 , denotes ,
T26483 703-705 NN denotes 1X
T26485 706-714 NN denotes Enhancer
T26484 715-723 NN denotes solution
T26486 723-725 , denotes ,
T26487 725-728 CD denotes 1.5
T26488 729-731 NN denotes mM
T26489 732-737 NN denotes MgSO4
T26490 737-739 , denotes ,
T26491 739-743 CD denotes 0.17
T26492 744-746 NN denotes mM
T26493 747-752 NNS denotes dNTPs
T26494 752-754 , denotes ,
T26495 754-755 CD denotes 1
T26496 756-758 NN denotes μM
T26498 759-763 DT denotes each
T26497 764-770 NN denotes primer
T26499 770-772 , denotes ,
T26500 772-775 CD denotes 0.1
T26501 776-780 NN denotes unit
T26502 781-783 IN denotes of
T26503 784-792 NN denotes Platinum
T26504 793-796 NN denotes Taq
T26505 797-798 -LRB- denotes (
T26506 798-808 NNP denotes Invitrogen
T26507 808-809 -RRB- denotes )
T26508 810-813 CC denotes and
T26509 814-827 RB denotes approximately
T26510 828-830 CD denotes 25
T26511 831-833 NN denotes ng
T26512 834-836 IN denotes of
T26513 837-841 NN denotes cDNA
T26514 841-842 . denotes .
T26515 842-1023 sentence denotes Reactions were incubated for 5 min at 95°C, then cycled for 45 s at 95°C, 45 s at 55°C, 1 min at 72°C for 35 cycles with a final 72°C extension for 10 min on a MJ Research PTC-200.
T26516 843-852 NNS denotes Reactions
T26518 853-857 VBD denotes were
T26517 858-867 VBN denotes incubated
T26519 868-871 IN denotes for
T26520 872-873 CD denotes 5
T26521 874-877 NN denotes min
T26522 878-880 IN denotes at
T26523 881-883 CD denotes 95
T26524 883-885 NN denotes °C
T26525 885-887 , denotes ,
T26526 887-891 RB denotes then
T26527 892-898 VBN denotes cycled
T26528 899-902 IN denotes for
T26529 903-905 CD denotes 45
T26530 906-907 NN denotes s
T26531 908-910 IN denotes at
T26532 911-913 CD denotes 95
T26533 913-915 NN denotes °C
T26534 915-917 , denotes ,
T26535 917-919 CD denotes 45
T26536 920-921 NN denotes s
T26537 922-924 IN denotes at
T26538 925-927 CD denotes 55
T26539 927-929 NN denotes °C
T26540 929-931 , denotes ,
T26541 931-932 CD denotes 1
T26542 933-936 NN denotes min
T26543 937-939 IN denotes at
T26544 940-942 CD denotes 72
T26545 942-944 NN denotes °C
T26546 945-948 IN denotes for
T26547 949-951 CD denotes 35
T26548 952-958 NNS denotes cycles
T26549 959-963 IN denotes with
T26550 964-965 DT denotes a
T26552 966-971 JJ denotes final
T26553 972-974 CD denotes 72
T26554 974-976 NN denotes °C
T26551 977-986 NN denotes extension
T26555 987-990 IN denotes for
T26556 991-993 CD denotes 10
T26557 994-997 NN denotes min
T26558 998-1000 IN denotes on
T26559 1001-1002 DT denotes a
T26561 1003-1005 NNP denotes MJ
T26562 1006-1014 NNP denotes Research
T26560 1015-1018 NNP denotes PTC
T26563 1018-1019 HYPH denotes -
T26564 1019-1022 CD denotes 200
T26565 1022-1023 . denotes .
T26566 1023-1101 sentence denotes The products were visualized on 1.5% agarose gels containing 0.06 μg/ml EtBr.
T26567 1024-1027 DT denotes The
T26568 1028-1036 NNS denotes products
T26570 1037-1041 VBD denotes were
T26569 1042-1052 VBN denotes visualized
T26571 1053-1055 IN denotes on
T26572 1056-1059 CD denotes 1.5
T26573 1059-1060 NN denotes %
T26574 1061-1068 NN denotes agarose
T26575 1069-1073 NNS denotes gels
T26576 1074-1084 VBG denotes containing
T26577 1085-1089 CD denotes 0.06
T26578 1090-1092 NN denotes μg
T26580 1092-1093 SYM denotes /
T26581 1093-1095 NN denotes ml
T26579 1096-1100 NN denotes EtBr
T26582 1100-1101 . denotes .
T26583 1101-1212 sentence denotes Bands of the correct size were excised and incubated in 100 μl of sterile H2O at 80°C for 10 min to elute DNA.
T26584 1102-1107 NNS denotes Bands
T26586 1108-1110 IN denotes of
T26587 1111-1114 DT denotes the
T26589 1115-1122 JJ denotes correct
T26588 1123-1127 NN denotes size
T26590 1128-1132 VBD denotes were
T26585 1133-1140 VBN denotes excised
T26591 1141-1144 CC denotes and
T26592 1145-1154 VBN denotes incubated
T26593 1155-1157 IN denotes in
T26594 1158-1161 CD denotes 100
T26595 1162-1164 NN denotes μl
T26596 1165-1167 IN denotes of
T26597 1168-1175 JJ denotes sterile
T26598 1176-1179 NN denotes H2O
T26599 1180-1182 IN denotes at
T26600 1183-1185 CD denotes 80
T26601 1185-1187 NN denotes °C
T26602 1188-1191 IN denotes for
T26603 1192-1194 CD denotes 10
T26604 1195-1198 NN denotes min
T26605 1199-1201 TO denotes to
T26606 1202-1207 VB denotes elute
T26607 1208-1211 NN denotes DNA
T26608 1211-1212 . denotes .
T26609 1212-1387 sentence denotes Reamplification reactions consisted of 1X PCR buffer, 1.5 mM MgCl2, 0.17 mM dNTPs, 1 μM each primer, 0.1 unit of Taq (Promega) and 10 μl of eluate in a total volume of 50 μl.
T26610 1213-1228 NN denotes Reamplification
T26611 1229-1238 NNS denotes reactions
T26612 1239-1248 VBD denotes consisted
T26613 1249-1251 IN denotes of
T26614 1252-1254 NN denotes 1X
T26616 1255-1258 NN denotes PCR
T26615 1259-1265 NN denotes buffer
T26617 1265-1267 , denotes ,
T26618 1267-1270 CD denotes 1.5
T26619 1271-1273 NN denotes mM
T26620 1274-1279 NN denotes MgCl2
T26621 1279-1281 , denotes ,
T26622 1281-1285 CD denotes 0.17
T26623 1286-1288 NN denotes mM
T26624 1289-1294 NNS denotes dNTPs
T26625 1294-1296 , denotes ,
T26626 1296-1297 CD denotes 1
T26627 1298-1300 NN denotes μM
T26629 1301-1305 DT denotes each
T26628 1306-1312 NN denotes primer
T26630 1312-1314 , denotes ,
T26631 1314-1317 CD denotes 0.1
T26632 1318-1322 NN denotes unit
T26633 1323-1325 IN denotes of
T26634 1326-1329 NN denotes Taq
T26635 1330-1331 -LRB- denotes (
T26636 1331-1338 NNP denotes Promega
T26637 1338-1339 -RRB- denotes )
T26638 1340-1343 CC denotes and
T26639 1344-1346 CD denotes 10
T26640 1347-1349 NN denotes μl
T26641 1350-1352 IN denotes of
T26642 1353-1359 NN denotes eluate
T26643 1360-1362 IN denotes in
T26644 1363-1364 DT denotes a
T26646 1365-1370 JJ denotes total
T26645 1371-1377 NN denotes volume
T26647 1378-1380 IN denotes of
T26648 1381-1383 CD denotes 50
T26649 1384-1386 NN denotes μl
T26650 1386-1387 . denotes .
T26651 1387-1456 sentence denotes The same cycling parameters were used for reamplification reactions.
T26652 1388-1391 DT denotes The
T26654 1392-1396 JJ denotes same
T26655 1397-1404 NN denotes cycling
T26653 1405-1415 NNS denotes parameters
T26657 1416-1420 VBD denotes were
T26656 1421-1425 VBN denotes used
T26658 1426-1429 IN denotes for
T26659 1430-1445 NN denotes reamplification
T26660 1446-1455 NNS denotes reactions
T26661 1455-1456 . denotes .
T26662 1456-1599 sentence denotes Products were visualized on 1% 0.5X TBE agarose gels containing 0.06 μg/ml EtBr, excised and purfied using PCR purification columns (Promega).
T26663 1457-1465 NNS denotes Products
T26665 1466-1470 VBD denotes were
T26664 1471-1481 VBN denotes visualized
T26666 1482-1484 IN denotes on
T26667 1485-1486 CD denotes 1
T26668 1486-1487 NN denotes %
T26670 1488-1492 NN denotes 0.5X
T26671 1493-1496 NN denotes TBE
T26672 1497-1504 NN denotes agarose
T26669 1505-1509 NNS denotes gels
T26673 1510-1520 VBG denotes containing
T26674 1521-1525 CD denotes 0.06
T26675 1526-1528 NN denotes μg
T26677 1528-1529 SYM denotes /
T26678 1529-1531 NN denotes ml
T26676 1532-1536 NN denotes EtBr
T26679 1536-1538 , denotes ,
T26680 1538-1545 VBN denotes excised
T26681 1546-1549 CC denotes and
T26682 1550-1557 VBN denotes purfied
T26683 1558-1563 VBG denotes using
T26684 1564-1567 NN denotes PCR
T26685 1568-1580 NN denotes purification
T26686 1581-1588 NNS denotes columns
T26687 1589-1590 -LRB- denotes (
T26688 1590-1597 NNP denotes Promega
T26689 1597-1598 -RRB- denotes )
T26690 1598-1599 . denotes .
T26691 1599-1756 sentence denotes To quantity each fragment 1 μl of each purified product was run on a 1.0% agarose gel containing 0.06 μg/ml EtBr, along with a DNA mass ladder (Invitrogen).
T26692 1600-1602 TO denotes To
T26693 1603-1611 VB denotes quantity
T26695 1612-1616 DT denotes each
T26696 1617-1625 NN denotes fragment
T26697 1626-1627 CD denotes 1
T26698 1628-1630 NN denotes μl
T26699 1631-1633 IN denotes of
T26700 1634-1638 DT denotes each
T26702 1639-1647 VBN denotes purified
T26701 1648-1655 NN denotes product
T26703 1656-1659 VBD denotes was
T26694 1660-1663 VBN denotes run
T26704 1664-1666 IN denotes on
T26705 1667-1668 DT denotes a
T26707 1669-1672 CD denotes 1.0
T26708 1672-1673 NN denotes %
T26709 1674-1681 NN denotes agarose
T26706 1682-1685 NN denotes gel
T26710 1686-1696 VBG denotes containing
T26711 1697-1701 CD denotes 0.06
T26712 1702-1704 NN denotes μg
T26714 1704-1705 SYM denotes /
T26715 1705-1707 NN denotes ml
T26713 1708-1712 NN denotes EtBr
T26716 1712-1714 , denotes ,
T26717 1714-1719 IN denotes along
T26718 1720-1724 IN denotes with
T26719 1725-1726 DT denotes a
T26721 1727-1730 NN denotes DNA
T26722 1731-1735 NN denotes mass
T26720 1736-1742 NN denotes ladder
T26723 1743-1744 -LRB- denotes (
T26724 1744-1754 NNP denotes Invitrogen
T26725 1754-1755 -RRB- denotes )
T26726 1755-1756 . denotes .
T26727 1756-1846 sentence denotes In a 96 well plate, 15 ng of each PCR product was added to 5 pM of primer for sequencing.
T26728 1757-1759 IN denotes In
T26730 1760-1761 DT denotes a
T26732 1762-1764 CD denotes 96
T26733 1765-1769 NN denotes well
T26731 1770-1775 NN denotes plate
T26734 1775-1777 , denotes ,
T26735 1777-1779 CD denotes 15
T26736 1780-1782 NN denotes ng
T26737 1783-1785 IN denotes of
T26738 1786-1790 DT denotes each
T26740 1791-1794 NN denotes PCR
T26739 1795-1802 NN denotes product
T26741 1803-1806 VBD denotes was
T26729 1807-1812 VBN denotes added
T26742 1813-1815 IN denotes to
T26743 1816-1817 CD denotes 5
T26744 1818-1820 NN denotes pM
T26745 1821-1823 IN denotes of
T26746 1824-1830 NN denotes primer
T26747 1831-1834 IN denotes for
T26748 1835-1845 NN denotes sequencing
T26749 1845-1846 . denotes .
T26750 1846-2018 sentence denotes The College of Agriculture and Environmental Sciences (CAES) Genomics Facility at the University of California, Davis, performed bidirectional sequencing of each amplicon.
T26751 1847-1850 DT denotes The
T26752 1851-1858 NNP denotes College
T26754 1859-1861 IN denotes of
T26755 1862-1873 NNP denotes Agriculture
T26757 1874-1877 CC denotes and
T26758 1878-1891 NNP denotes Environmental
T26756 1892-1900 NNPS denotes Sciences
T26759 1901-1902 -LRB- denotes (
T26760 1902-1906 NNP denotes CAES
T26761 1906-1907 -RRB- denotes )
T26762 1908-1916 NNP denotes Genomics
T26753 1917-1925 NN denotes Facility
T26764 1926-1928 IN denotes at
T26765 1929-1932 DT denotes the
T26766 1933-1943 NNP denotes University
T26767 1944-1946 IN denotes of
T26768 1947-1957 NNP denotes California
T26769 1957-1959 , denotes ,
T26770 1959-1964 NNP denotes Davis
T26771 1964-1966 , denotes ,
T26763 1966-1975 VBD denotes performed
T26772 1976-1989 JJ denotes bidirectional
T26773 1990-2000 NN denotes sequencing
T26774 2001-2003 IN denotes of
T26775 2004-2008 DT denotes each
T26776 2009-2017 NN denotes amplicon
T26777 2017-2018 . denotes .
R8043 T26350 T26351 compound Candidate,gene
R8044 T26351 T26352 compound gene,sequencing
R8045 T26354 T26355 compound PCR,amplicons
R8046 T26355 T26356 nsubjpass amplicons,amplified
R8047 T26357 T26355 acl covering,amplicons
R8048 T26358 T26359 det the,sequence
R8049 T26359 T26357 dobj sequence,covering
R8050 T26360 T26359 compound coding,sequence
R8051 T26361 T26355 cc and,amplicons
R8052 T26362 T26363 amod partial,regions
R8053 T26363 T26355 conj regions,amplicons
R8054 T26364 T26363 nummod 5,regions
R8055 T26365 T26364 punct ',5
R8056 T26366 T26364 cc and,5
R8057 T26367 T26364 conj 3,5
R8058 T26368 T26367 punct ',3
R8059 T26369 T26363 amod untranslated,regions
R8060 T26370 T26363 prep for,regions
R8061 T26371 T26372 det each,gene
R8062 T26372 T26370 pobj gene,for
R8063 T26373 T26372 amod selected,gene
R8064 T26374 T26372 compound candidate,gene
R8065 T26375 T26356 auxpass were,amplified
R8066 T26376 T26356 punct ", ",amplified
R8067 T26377 T26356 conj purified,amplified
R8068 T26378 T26377 cc and,purified
R8069 T26379 T26377 conj sequenced,purified
R8070 T26380 T26379 prep from,sequenced
R8071 T26381 T26382 det the,strain
R8072 T26382 T26380 pobj strain,from
R8073 T26383 T26382 compound CAST,strain
R8074 T26384 T26379 advcl using,sequenced
R8075 T26385 T26384 dobj protocols,using
R8076 T26386 T26385 acl outlined,protocols
R8077 T26387 T26386 prep in,outlined
R8078 T26388 T26389 punct [,57
R8079 T26389 T26387 pobj 57,in
R8080 T26390 T26389 punct ],57
R8081 T26391 T26385 prep with,protocols
R8082 T26392 T26393 amod slight,modifications
R8083 T26393 T26391 pobj modifications,with
R8084 T26394 T26356 punct .,amplified
R8085 T26396 T26397 amod Total,RNA
R8086 T26397 T26398 nsubjpass RNA,isolated
R8087 T26399 T26397 prep from,RNA
R8088 T26400 T26399 pobj brain,from
R8089 T26401 T26400 punct ", ",brain
R8090 T26402 T26400 conj liver,brain
R8091 T26403 T26402 punct ", ",liver
R8092 T26404 T26402 conj spleen,liver
R8093 T26405 T26404 punct ", ",spleen
R8094 T26406 T26404 conj lung,spleen
R8095 T26407 T26406 cc and,lung
R8096 T26408 T26406 conj testis,lung
R8097 T26409 T26398 auxpass was,isolated
R8098 T26410 T26398 prep from,isolated
R8099 T26411 T26412 det an,mouse
R8100 T26412 T26410 pobj mouse,from
R8101 T26413 T26412 amod adult,mouse
R8102 T26414 T26412 nmod CAST,mouse
R8103 T26415 T26412 amod male,mouse
R8104 T26416 T26398 advcl using,isolated
R8105 T26417 T26416 dobj Trizol,using
R8106 T26418 T26419 punct (,Ambion
R8107 T26419 T26417 parataxis Ambion,Trizol
R8108 T26420 T26419 punct ),Ambion
R8109 T26421 T26398 punct .,isolated
R8110 T26423 T26424 nsubjpass cDNA,produced
R8111 T26425 T26424 auxpass was,produced
R8112 T26426 T26424 prep from,produced
R8113 T26427 T26428 amod total,RNA
R8114 T26428 T26426 pobj RNA,from
R8115 T26429 T26424 advcl using,produced
R8116 T26430 T26431 amod standard,procedures
R8117 T26431 T26429 dobj procedures,using
R8118 T26432 T26424 punct .,produced
R8119 T26434 T26435 compound PCR,sets
R8120 T26435 T26437 nsubjpass sets,designed
R8121 T26436 T26435 compound primer,sets
R8122 T26438 T26435 prep for,sets
R8123 T26439 T26440 det each,gene
R8124 T26440 T26438 pobj gene,for
R8125 T26441 T26437 auxpass were,designed
R8126 T26442 T26437 advcl using,designed
R8127 T26443 T26442 dobj Primer3,using
R8128 T26444 T26445 punct [,58
R8129 T26445 T26437 parataxis 58,designed
R8130 T26446 T26445 punct ],58
R8131 T26447 T26448 punct (,File
R8132 T26448 T26437 parataxis File,designed
R8133 T26449 T26448 amod Additional,File
R8134 T26450 T26448 nummod 5,File
R8135 T26451 T26448 punct ),File
R8136 T26452 T26437 punct .,designed
R8137 T26454 T26455 det The,amplification
R8138 T26455 T26457 nsubjpass amplification,performed
R8139 T26456 T26455 amod initial,amplification
R8140 T26458 T26457 auxpass was,performed
R8141 T26459 T26457 prep in,performed
R8142 T26460 T26461 nummod 10,μl
R8143 T26461 T26462 compound μl,reactions
R8144 T26462 T26459 pobj reactions,in
R8145 T26463 T26462 compound PCR,reactions
R8146 T26464 T26457 advcl using,performed
R8147 T26465 T26466 det the,system
R8148 T26466 T26464 dobj system,using
R8149 T26467 T26466 compound Invitrogen,system
R8150 T26468 T26466 compound Platinum,system
R8151 T26469 T26470 compound TaqPCRx,amplification
R8152 T26470 T26466 compound amplification,system
R8153 T26471 T26472 punct (,Invitrogen
R8154 T26472 T26466 parataxis Invitrogen,system
R8155 T26473 T26472 punct ),Invitrogen
R8156 T26474 T26457 punct .,performed
R8157 T26476 T26477 det The,reactions
R8158 T26477 T26478 nsubj reactions,contained
R8159 T26479 T26480 compound 1X,buffer
R8160 T26480 T26478 dobj buffer,contained
R8161 T26481 T26480 compound PCR,buffer
R8162 T26482 T26480 punct ", ",buffer
R8163 T26483 T26484 compound 1X,solution
R8164 T26484 T26480 conj solution,buffer
R8165 T26485 T26484 compound Enhancer,solution
R8166 T26486 T26484 punct ", ",solution
R8167 T26487 T26488 nummod 1.5,mM
R8168 T26488 T26489 compound mM,MgSO4
R8169 T26489 T26484 conj MgSO4,solution
R8170 T26490 T26489 punct ", ",MgSO4
R8171 T26491 T26492 nummod 0.17,mM
R8172 T26492 T26493 compound mM,dNTPs
R8173 T26493 T26489 conj dNTPs,MgSO4
R8174 T26494 T26493 punct ", ",dNTPs
R8175 T26495 T26496 nummod 1,μM
R8176 T26496 T26497 nmod μM,primer
R8177 T26497 T26493 conj primer,dNTPs
R8178 T26498 T26497 det each,primer
R8179 T26499 T26497 punct ", ",primer
R8180 T26500 T26501 nummod 0.1,unit
R8181 T26501 T26497 conj unit,primer
R8182 T26502 T26501 prep of,unit
R8183 T26503 T26504 compound Platinum,Taq
R8184 T26504 T26502 pobj Taq,of
R8185 T26505 T26506 punct (,Invitrogen
R8186 T26506 T26504 parataxis Invitrogen,Taq
R8187 T26507 T26506 punct ),Invitrogen
R8188 T26508 T26501 cc and,unit
R8189 T26509 T26510 advmod approximately,25
R8190 T26510 T26511 nummod 25,ng
R8191 T26511 T26501 conj ng,unit
R8192 T26512 T26511 prep of,ng
R8193 T26513 T26512 pobj cDNA,of
R8194 T26514 T26478 punct .,contained
R8195 T26516 T26517 nsubjpass Reactions,incubated
R8196 T26518 T26517 auxpass were,incubated
R8197 T26519 T26517 prep for,incubated
R8198 T26520 T26521 nummod 5,min
R8199 T26521 T26519 pobj min,for
R8200 T26522 T26517 prep at,incubated
R8201 T26523 T26524 nummod 95,°C
R8202 T26524 T26522 pobj °C,at
R8203 T26525 T26517 punct ", ",incubated
R8204 T26526 T26527 advmod then,cycled
R8205 T26527 T26517 dep cycled,incubated
R8206 T26528 T26527 prep for,cycled
R8207 T26529 T26530 nummod 45,s
R8208 T26530 T26528 pobj s,for
R8209 T26531 T26530 prep at,s
R8210 T26532 T26533 nummod 95,°C
R8211 T26533 T26531 pobj °C,at
R8212 T26534 T26530 punct ", ",s
R8213 T26535 T26536 nummod 45,s
R8214 T26536 T26530 appos s,s
R8215 T26537 T26536 prep at,s
R8216 T26538 T26539 nummod 55,°C
R8217 T26539 T26537 pobj °C,at
R8218 T26540 T26530 punct ", ",s
R8219 T26541 T26542 nummod 1,min
R8220 T26542 T26530 appos min,s
R8221 T26543 T26542 prep at,min
R8222 T26544 T26545 nummod 72,°C
R8223 T26545 T26543 pobj °C,at
R8224 T26546 T26542 prep for,min
R8225 T26547 T26548 nummod 35,cycles
R8226 T26548 T26546 pobj cycles,for
R8227 T26549 T26527 prep with,cycled
R8228 T26550 T26551 det a,extension
R8229 T26551 T26549 pobj extension,with
R8230 T26552 T26551 amod final,extension
R8231 T26553 T26554 nummod 72,°C
R8232 T26554 T26551 compound °C,extension
R8233 T26555 T26551 prep for,extension
R8234 T26556 T26557 nummod 10,min
R8235 T26557 T26555 pobj min,for
R8236 T26558 T26551 prep on,extension
R8237 T26559 T26560 det a,PTC
R8238 T26560 T26558 pobj PTC,on
R8239 T26561 T26562 compound MJ,Research
R8240 T26562 T26560 compound Research,PTC
R8241 T26563 T26560 punct -,PTC
R8242 T26564 T26560 nummod 200,PTC
R8243 T26565 T26517 punct .,incubated
R8244 T26567 T26568 det The,products
R8245 T26568 T26569 nsubjpass products,visualized
R8246 T26570 T26569 auxpass were,visualized
R8247 T26571 T26569 prep on,visualized
R8248 T26572 T26573 nummod 1.5,%
R8249 T26573 T26574 compound %,agarose
R8250 T26574 T26575 compound agarose,gels
R8251 T26575 T26571 pobj gels,on
R8252 T26576 T26575 acl containing,gels
R8253 T26577 T26578 nummod 0.06,μg
R8254 T26578 T26579 nmod μg,EtBr
R8255 T26579 T26576 dobj EtBr,containing
R8256 T26580 T26581 punct /,ml
R8257 T26581 T26578 prep ml,μg
R8258 T26582 T26569 punct .,visualized
R8259 T26584 T26585 nsubjpass Bands,excised
R8260 T26586 T26584 prep of,Bands
R8261 T26587 T26588 det the,size
R8262 T26588 T26586 pobj size,of
R8263 T26589 T26588 amod correct,size
R8264 T26590 T26585 auxpass were,excised
R8265 T26591 T26585 cc and,excised
R8266 T26592 T26585 conj incubated,excised
R8267 T26593 T26592 prep in,incubated
R8268 T26594 T26595 nummod 100,μl
R8269 T26595 T26593 pobj μl,in
R8270 T26596 T26595 prep of,μl
R8271 T26597 T26598 amod sterile,H2O
R8272 T26598 T26596 pobj H2O,of
R8273 T26599 T26598 prep at,H2O
R8274 T26600 T26601 nummod 80,°C
R8275 T26601 T26599 pobj °C,at
R8276 T26602 T26592 prep for,incubated
R8277 T26603 T26604 nummod 10,min
R8278 T26604 T26602 pobj min,for
R8279 T26605 T26606 aux to,elute
R8280 T26606 T26592 advcl elute,incubated
R8281 T26607 T26606 dobj DNA,elute
R8282 T26608 T26585 punct .,excised
R8283 T26610 T26611 compound Reamplification,reactions
R8284 T26611 T26612 nsubj reactions,consisted
R8285 T26613 T26612 prep of,consisted
R8286 T26614 T26615 compound 1X,buffer
R8287 T26615 T26613 pobj buffer,of
R8288 T26616 T26615 compound PCR,buffer
R8289 T26617 T26615 punct ", ",buffer
R8290 T26618 T26619 nummod 1.5,mM
R8291 T26619 T26620 compound mM,MgCl2
R8292 T26620 T26615 conj MgCl2,buffer
R8293 T26621 T26620 punct ", ",MgCl2
R8294 T26622 T26623 nummod 0.17,mM
R8295 T26623 T26624 compound mM,dNTPs
R8296 T26624 T26620 conj dNTPs,MgCl2
R8297 T26625 T26624 punct ", ",dNTPs
R8298 T26626 T26627 nummod 1,μM
R8299 T26627 T26628 nmod μM,primer
R8300 T26628 T26624 conj primer,dNTPs
R8301 T26629 T26628 det each,primer
R8302 T26630 T26628 punct ", ",primer
R8303 T26631 T26632 nummod 0.1,unit
R8304 T26632 T26628 conj unit,primer
R8305 T26633 T26632 prep of,unit
R8306 T26634 T26633 pobj Taq,of
R8307 T26635 T26636 punct (,Promega
R8308 T26636 T26634 parataxis Promega,Taq
R8309 T26637 T26636 punct ),Promega
R8310 T26638 T26632 cc and,unit
R8311 T26639 T26640 nummod 10,μl
R8312 T26640 T26632 conj μl,unit
R8313 T26641 T26640 prep of,μl
R8314 T26642 T26641 pobj eluate,of
R8315 T26643 T26615 prep in,buffer
R8316 T26644 T26645 det a,volume
R8317 T26645 T26643 pobj volume,in
R8318 T26646 T26645 amod total,volume
R8319 T26647 T26645 prep of,volume
R8320 T26648 T26649 nummod 50,μl
R8321 T26649 T26647 pobj μl,of
R8322 T26650 T26612 punct .,consisted
R8323 T26652 T26653 det The,parameters
R8324 T26653 T26656 nsubjpass parameters,used
R8325 T26654 T26653 amod same,parameters
R8326 T26655 T26653 compound cycling,parameters
R8327 T26657 T26656 auxpass were,used
R8328 T26658 T26656 prep for,used
R8329 T26659 T26660 compound reamplification,reactions
R8330 T26660 T26658 pobj reactions,for
R8331 T26661 T26656 punct .,used
R8332 T26663 T26664 nsubjpass Products,visualized
R8333 T26665 T26664 auxpass were,visualized
R8334 T26666 T26664 prep on,visualized
R8335 T26667 T26668 nummod 1,%
R8336 T26668 T26669 compound %,gels
R8337 T26669 T26666 pobj gels,on
R8338 T26670 T26669 compound 0.5X,gels
R8339 T26671 T26669 compound TBE,gels
R8340 T26672 T26669 compound agarose,gels
R8341 T26673 T26669 acl containing,gels
R8342 T26674 T26675 nummod 0.06,μg
R8343 T26675 T26676 nmod μg,EtBr
R8344 T26676 T26673 dobj EtBr,containing
R8345 T26677 T26678 punct /,ml
R8346 T26678 T26675 prep ml,μg
R8347 T26679 T26664 punct ", ",visualized
R8348 T26680 T26664 conj excised,visualized
R8349 T26681 T26680 cc and,excised
R8350 T26682 T26680 conj purfied,excised
R8351 T26683 T26682 advcl using,purfied
R8352 T26684 T26685 compound PCR,purification
R8353 T26685 T26686 compound purification,columns
R8354 T26686 T26683 dobj columns,using
R8355 T26687 T26688 punct (,Promega
R8356 T26688 T26686 parataxis Promega,columns
R8357 T26689 T26688 punct ),Promega
R8358 T26690 T26664 punct .,visualized
R8359 T26692 T26693 aux To,quantity
R8360 T26693 T26694 advcl quantity,run
R8361 T26695 T26696 det each,fragment
R8362 T26696 T26693 dobj fragment,quantity
R8363 T26697 T26698 nummod 1,μl
R8364 T26698 T26694 nsubjpass μl,run
R8365 T26699 T26698 prep of,μl
R8366 T26700 T26701 det each,product
R8367 T26701 T26699 pobj product,of
R8368 T26702 T26701 amod purified,product
R8369 T26703 T26694 auxpass was,run
R8370 T26704 T26694 prep on,run
R8371 T26705 T26706 det a,gel
R8372 T26706 T26704 pobj gel,on
R8373 T26707 T26708 nummod 1.0,%
R8374 T26708 T26706 compound %,gel
R8375 T26709 T26706 compound agarose,gel
R8376 T26710 T26706 acl containing,gel
R8377 T26711 T26712 nummod 0.06,μg
R8378 T26712 T26713 nmod μg,EtBr
R8379 T26713 T26710 dobj EtBr,containing
R8380 T26714 T26715 punct /,ml
R8381 T26715 T26712 prep ml,μg
R8382 T26716 T26706 punct ", ",gel
R8383 T26717 T26706 prep along,gel
R8384 T26718 T26717 prep with,along
R8385 T26719 T26720 det a,ladder
R8386 T26720 T26718 pobj ladder,with
R8387 T26721 T26720 compound DNA,ladder
R8388 T26722 T26720 compound mass,ladder
R8389 T26723 T26724 punct (,Invitrogen
R8390 T26724 T26720 parataxis Invitrogen,ladder
R8391 T26725 T26724 punct ),Invitrogen
R8392 T26726 T26694 punct .,run
R8393 T26728 T26729 prep In,added
R8394 T26730 T26731 det a,plate
R8395 T26731 T26728 pobj plate,In
R8396 T26732 T26733 nummod 96,well
R8397 T26733 T26731 compound well,plate
R8398 T26734 T26729 punct ", ",added
R8399 T26735 T26736 nummod 15,ng
R8400 T26736 T26729 nsubjpass ng,added
R8401 T26737 T26736 prep of,ng
R8402 T26738 T26739 det each,product
R8403 T26739 T26737 pobj product,of
R8404 T26740 T26739 compound PCR,product
R8405 T26741 T26729 auxpass was,added
R8406 T26742 T26729 prep to,added
R8407 T26743 T26744 nummod 5,pM
R8408 T26744 T26742 pobj pM,to
R8409 T26745 T26744 prep of,pM
R8410 T26746 T26745 pobj primer,of
R8411 T26747 T26729 prep for,added
R8412 T26748 T26747 pobj sequencing,for
R8413 T26749 T26729 punct .,added
R8414 T26751 T26752 det The,College
R8415 T26752 T26753 nmod College,Facility
R8416 T26753 T26763 nsubj Facility,performed
R8417 T26754 T26752 prep of,College
R8418 T26755 T26756 nmod Agriculture,Sciences
R8419 T26756 T26754 pobj Sciences,of
R8420 T26757 T26755 cc and,Agriculture
R8421 T26758 T26755 conj Environmental,Agriculture
R8422 T26759 T26752 punct (,College
R8423 T26760 T26752 appos CAES,College
R8424 T26761 T26753 punct ),Facility
R8425 T26762 T26753 compound Genomics,Facility
R8426 T26764 T26753 prep at,Facility
R8427 T26765 T26766 det the,University
R8428 T26766 T26764 pobj University,at
R8429 T26767 T26766 prep of,University
R8430 T26768 T26767 pobj California,of
R8431 T26769 T26766 punct ", ",University
R8432 T26770 T26766 npadvmod Davis,University
R8433 T26771 T26763 punct ", ",performed
R8434 T26772 T26773 amod bidirectional,sequencing
R8435 T26773 T26763 dobj sequencing,performed
R8436 T26774 T26773 prep of,sequencing
R8437 T26775 T26776 det each,amplicon
R8438 T26776 T26774 pobj amplicon,of
R8439 T26777 T26763 punct .,performed