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of ES cells bearing the Atrx flox allele.\nBriefly, the targeting vector (shown in Figure 2A) places a loxP site within intron 18 and a loxP-flanked MC1neopA selection cassette in intron 17 of the Atrx gene. 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{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T11149","span":{"begin":0,"end":13},"obj":"_FRAGMENT"},{"id":"T11150","span":{"begin":17,"end":22},"obj":"GO:0048468"},{"id":"T11151","span":{"begin":14,"end":22},"obj":"CL:0002322"},{"id":"T11152","span":{"begin":17,"end":22},"obj":"CL_GO_EXT:cell"},{"id":"T11153","span":{"begin":35,"end":39},"obj":"PR_EXT:000004503"},{"id":"T11154","span":{"begin":40,"end":44},"obj":"SO:0000359"},{"id":"T11155","span":{"begin":45,"end":51},"obj":"SO_EXT:0001023"},{"id":"T11156","span":{"begin":66,"end":82},"obj":"SO_EXT:0001644"},{"id":"T11157","span":{"begin":113,"end":122},"obj":"SO_EXT:0000346"},{"id":"T11158","span":{"begin":130,"end":136},"obj":"SO_EXT:0000188"},{"id":"T11159","span":{"begin":146,"end":158},"obj":"SO:0000359"},{"id":"T11160","span":{"begin":178,"end":186},"obj":"SO_EXT:0005853"},{"id":"T11161","span":{"begin":190,"end":196},"obj":"SO_EXT:0000188"},{"id":"T11162","span":{"begin":207,"end":211},"obj":"PR_EXT:000004503"},{"id":"T11163","span":{"begin":212,"end":216},"obj":"SO_EXT:0000704"},{"id":"T11164","span":{"begin":258,"end":267},"obj":"SO_EXT:engineered_biological_sequence"},{"id":"T11165","span":{"begin":289,"end":299},"obj":"SO_EXT:sequence_linearization"},{"id":"T11166","span":{"begin":300,"end":307},"obj":"SO_EXT:0000155"},{"id":"T11167","span":{"begin":357,"end":365},"obj":"CL:0002322"},{"id":"T11168","span":{"begin":360,"end":365},"obj":"CL_GO_EXT:cell"},{"id":"T11169","span":{"begin":393,"end":397},"obj":"CHEBI:42768"},{"id":"T11170","span":{"begin":402,"end":413},"obj":"CHEBI:465284"},{"id":"T11171","span":{"begin":508,"end":511},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T11172","span":{"begin":516,"end":529},"obj":"GO:0097617"},{"id":"T11173","span":{"begin":540,"end":545},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T11174","span":{"begin":562,"end":569},"obj":"SO_EXT:0000112"},{"id":"T11175","span":{"begin":600,"end":605},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T11176","span":{"begin":614,"end":615},"obj":"CHEBI_SO_EXT:base"},{"id":"T11177","span":{"begin":663,"end":666},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T11178","span":{"begin":691,"end":697},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T11179","span":{"begin":760,"end":773},"obj":"GO:0097617"},{"id":"T11180","span":{"begin":781,"end":786},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T11181","span":{"begin":799,"end":805},"obj":"SO_EXT:0000188"},{"id":"T11182","span":{"begin":812,"end":823},"obj":"SO_EXT:0000006"},{"id":"T11183","span":{"begin":839,"end":846},"obj":"SO_EXT:0000112"},{"id":"T11184","span":{"begin":886,"end":895},"obj":"SO_EXT:0000346"},{"id":"T11185","span":{"begin":903,"end":909},"obj":"SO_EXT:0000188"},{"id":"T11186","span":{"begin":942,"end":954},"obj":"GO:0035825"},{"id":"T11187","span":{"begin":982,"end":991},"obj":"SO_EXT:biological_sequence"},{"id":"T11188","span":{"begin":995,"end":1002},"obj":"SO_EXT:0000112"}],"relations":[{"id":"R6940","pred":"_lexicallyChainedTo","subj":"T11150","obj":"T11149"}],"text":"Generation of ES cells bearing the Atrx flox allele.\nBriefly, the targeting vector (shown in Figure 2A) places a loxP site within intron 18 and a loxP-flanked MC1neopA selection cassette in intron 17 of the Atrx gene. A detailed description of the targeting construct is provided in [20]. Linearised plasmid (150 μg) was electroporated into 1 × 108 E14Tg2a ES cells, and colonies resistant to G418 and ganciclovir were isolated. Homologous targeting events were identified by Southern blot of EcoRI-digested DNA and hybridisation with a 5′ probe (generated with primers PPS1.20 and PPS1.27) and a 3′ probe (a 0.9-kb HaeIII fragment) as shown in Figure 2A and 2B. DNA from correctly targeted clones was also digested with SacI and analysed by Southern blot and hybridisation with a probe from within intron 17 (a PCR product generated with primers PPS1.15 and Xnp46) to confirm that the loxP site within intron 18 had been included within the crossed-over region (Figure 2A and 2C). Sequences of primers are shown in Table S1."}

{"project":"2_test","denotations":[{"id":"16628246-15668733-85799784","span":{"begin":284,"end":286},"obj":"15668733"},{"id":"T64563","span":{"begin":284,"end":286},"obj":"15668733"}],"text":"Generation of ES cells bearing the Atrx flox allele.\nBriefly, the targeting vector (shown in Figure 2A) places a loxP site within intron 18 and a loxP-flanked MC1neopA selection cassette in intron 17 of the Atrx gene. A detailed description of the targeting construct is provided in [20]. Linearised plasmid (150 μg) was electroporated into 1 × 108 E14Tg2a ES cells, and colonies resistant to G418 and ganciclovir were isolated. Homologous targeting events were identified by Southern blot of EcoRI-digested DNA and hybridisation with a 5′ probe (generated with primers PPS1.20 and PPS1.27) and a 3′ probe (a 0.9-kb HaeIII fragment) as shown in Figure 2A and 2B. DNA from correctly targeted clones was also digested with SacI and analysed by Southern blot and hybridisation with a probe from within intron 17 (a PCR product generated with primers PPS1.15 and Xnp46) to confirm that the loxP site within intron 18 had been included within the crossed-over region (Figure 2A and 2C). Sequences of primers are shown in Table S1."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T11121","span":{"begin":0,"end":13},"obj":"_FRAGMENT"},{"id":"T11122","span":{"begin":17,"end":22},"obj":"GO:0048468"},{"id":"T11123","span":{"begin":14,"end":22},"obj":"CL:0002322"},{"id":"T11124","span":{"begin":35,"end":39},"obj":"PR:000004503"},{"id":"T11125","span":{"begin":40,"end":44},"obj":"SO:0000359"},{"id":"T11126","span":{"begin":45,"end":51},"obj":"SO:0001023"},{"id":"T11127","span":{"begin":66,"end":82},"obj":"SO:0001644"},{"id":"T11128","span":{"begin":113,"end":122},"obj":"SO:0000346"},{"id":"T11129","span":{"begin":130,"end":136},"obj":"SO:0000188"},{"id":"T11130","span":{"begin":146,"end":158},"obj":"SO:0000359"},{"id":"T11131","span":{"begin":178,"end":186},"obj":"SO:0005853"},{"id":"T11132","span":{"begin":190,"end":196},"obj":"SO:0000188"},{"id":"T11133","span":{"begin":207,"end":211},"obj":"PR:000004503"},{"id":"T11134","span":{"begin":212,"end":216},"obj":"SO:0000704"},{"id":"T11135","span":{"begin":300,"end":307},"obj":"SO:0000155"},{"id":"T11136","span":{"begin":357,"end":365},"obj":"CL:0002322"},{"id":"T11137","span":{"begin":393,"end":397},"obj":"CHEBI:42768"},{"id":"T11138","span":{"begin":402,"end":413},"obj":"CHEBI:465284"},{"id":"T11139","span":{"begin":516,"end":529},"obj":"GO:0097617"},{"id":"T11140","span":{"begin":562,"end":569},"obj":"SO:0000112"},{"id":"T11141","span":{"begin":760,"end":773},"obj":"GO:0097617"},{"id":"T11142","span":{"begin":799,"end":805},"obj":"SO:0000188"},{"id":"T11143","span":{"begin":812,"end":823},"obj":"SO:0000006"},{"id":"T11144","span":{"begin":839,"end":846},"obj":"SO:0000112"},{"id":"T11145","span":{"begin":886,"end":895},"obj":"SO:0000346"},{"id":"T11146","span":{"begin":903,"end":909},"obj":"SO:0000188"},{"id":"T11147","span":{"begin":942,"end":954},"obj":"GO:0035825"},{"id":"T11148","span":{"begin":995,"end":1002},"obj":"SO:0000112"}],"relations":[{"id":"R6939","pred":"_lexicallyChainedTo","subj":"T11122","obj":"T11121"}],"text":"Generation of ES cells bearing the Atrx flox allele.\nBriefly, the targeting vector (shown in Figure 2A) places a loxP site within intron 18 and a loxP-flanked MC1neopA selection cassette in intron 17 of the Atrx gene. A detailed description of the targeting construct is provided in [20]. Linearised plasmid (150 μg) was electroporated into 1 × 108 E14Tg2a ES cells, and colonies resistant to G418 and ganciclovir were isolated. Homologous targeting events were identified by Southern blot of EcoRI-digested DNA and hybridisation with a 5′ probe (generated with primers PPS1.20 and PPS1.27) and a 3′ probe (a 0.9-kb HaeIII fragment) as shown in Figure 2A and 2B. DNA from correctly targeted clones was also digested with SacI and analysed by Southern blot and hybridisation with a probe from within intron 17 (a PCR product generated with primers PPS1.15 and Xnp46) to confirm that the loxP site within intron 18 had been included within the crossed-over region (Figure 2A and 2C). Sequences of primers are shown in Table S1."}