F12
Primary Info
Mutation
Mouse studies
DNA sequencing


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I. DNA amplification
Kit
GE:illustra GenomiPhi V2 DNA Amplification Kit

II. PCR
Primer details
Exon No. Forward sequence Reverse sequence Expected PCR
product size (bp)		 Genomic regions amplified by ePCR with
the primer
1 TGACCAAGGAAGTTGCTCCAC GGCCTCCTAGTCACCTGGAC 322bp chr5:176836361-176836682
2 TTTGGGTCTTGGGAAGAAATC GCTCACGATCACTCTAGTGCC 318bp chr5:176835908-176836225
3 CTTGATCTCCTGACCTCGTG CACCCTGCCCTATCACAGTC 436bp chr5:176832853-176833288
4-5 GACTGTGATAGGGCAGGGTG ATTCTGTAGGCCCAGGGTTG 614bp chr5:176832259-176832872
6-7 GGTTGCTGGATACTCGGAGAC TCCTGCCTCCCTCTCATCTG 550bp chr5:176831708-176832257
8 AGGAGGGAGAGTGCGGAAAG CGTTGTCCGGGTTCCTGTAG 347bp chr5:176831401-176831747
9 ACCTACCGGAACGTGACTGC CGGGTCATCGAAGACAGACTC 570bp chr5:176830995-176831564
10 CCTAGGCTTCATGTCCCACTC AGAAGGTAGGGCACGGGTTC 528bp chr5:176830763-176831290
11-12 GAAGCTGGAACACGGGATTG GTAGGCACCCGGAACGATAC 562bp chr5:176830156-176830717
13 TGAATTAATGGCACCTCCTCC GCTTGGTCCTCACACACCAG 424bp chr5:176829423-176829846
14 CGTGCCAGGTGAGCTCTTAG GTGGAGAAGCTGTGAATCGC 490bp chr5:176829069-176829558

III. Sequencing reaction
Kit
BigDye® Terminator v3.1 Cycle Sequencing Kit
Primer details
Exon No. Forward sequence Reverse sequence Expected PCR
product size (bp)		 Genomic regions amplified by ePCR with
the primer
1 TGACCAAGGAAGTTGCTCCAC GGCCTCCTAGTCACCTGGAC 322bp chr5:176836361-176836682
2 TTTGGGTCTTGGGAAGAAATC GCTCACGATCACTCTAGTGCC 318bp chr5:176835908-176836225
3 CTTGATCTCCTGACCTCGTG CACCCTGCCCTATCACAGTC 436bp chr5:176832853-176833288
4-5 GACTGTGATAGGGCAGGGTG ATTCTGTAGGCCCAGGGTTG 614bp chr5:176832259-176832872
6-7 GGTTGCTGGATACTCGGAGAC TCCTGCCTCCCTCTCATCTG 550bp chr5:176831708-176832257
8 AGGAGGGAGAGTGCGGAAAG CGTTGTCCGGGTTCCTGTAG 347bp chr5:176831401-176831747
9 ACCTACCGGAACGTGACTGC CGGGTCATCGAAGACAGACTC 570bp chr5:176830995-176831564
10 CCTAGGCTTCATGTCCCACTC AGAAGGTAGGGCACGGGTTC 528bp chr5:176830763-176831290
11-12 GAAGCTGGAACACGGGATTG GTAGGCACCCGGAACGATAC 562bp chr5:176830156-176830717
13 TGAATTAATGGCACCTCCTCC GCTTGGTCCTCACACACCAG 424bp chr5:176829423-176829846
14 CGTGCCAGGTGAGCTCTTAG GTGGAGAAGCTGTGAATCGC 490bp chr5:176829069-176829558

IV. Ethanol precipitation

V. Sequencing analysis