UNC13D
Primary Info
Mutation
Mouse studies
DNA sequencing


Click on each title for more information
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 CCCAGGCTCTTTGCATAATC GAGGTGGGGCCAGACAG 282bp chr17:73840216-73840497
2-3-4 TCAGAGGAGCCATTCCTGTC CCAGAATGCCCTTGGCCTGT 723bp chr17:73838953-73839675
5-6 CAGACACTGCAGCGGGTCA CCTGAGCCAGGACGACCT 708bp chr17:73838417-73839124
7-8-9 GAGGGAAGTTTGGCAGGGATAG GACAGGAGGTGGACCCCA chr17:73837179-?
10-11-12 GGGTCCACCTCCTGTCG TACACTTTGGGCTACCGGC chr17:?-73835853
13-14-15 GAGCCACTAGTTTTCAGCCTG GGTGGAACCATTCAGTGGTGC 809bp chr17:73832313-73833121
16-17-18 CCATTGCCCCAGCTGGTGA TGTGGTCCTGCACCCGCTT 619bp chr17:73831834-73832452
19-20 CCTCTTCTCCATGGCTTTCCG GGGATGTTCAGAGCGGGTTAC 593bp chr17:73831380-73831972
21-22 TCTGTGCCTGGTGATGGTAG GCTTCCTGAACTGTGCCC 607bp chr17:73830631-73831237
23-24 CCAACATGGTAAGGCCAGAG ACCAAGGCCTTCAATGACC 626bp chr17:73830081-73830706
25 CTCCAGGTCAGGGGAAGG CCTCAGAACGGATGCTGG 248bp chr17:73828956-73829203
26-27 AGAGCGTCTTTGCTTCCTCC TTATGTAGCCGACCCTGGAC 407bp chr17:73827094-73827500
28-29 AAACGGACATGTTACCCTTGAC CTCCTGCAGGGTGAGCGAA 491bp chr17:73826352-73826842
30 AGCATAGCTGCTGGGGTG CATCTCAGGGGAGCCCCTT 296bp chr17:73826005-73826300
31 CCAACACCAAGTGGGTGAAC GACCCTGGCATCACCTCTC 365bp chr17:73824783-73825147
32 GGAGCAGTGTTGTGTACCCC TCTGGGCACTCCGCATGCT 359bp chr17:73823906-73824264

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 CCCAGGCTCTTTGCATAATC ACCCGAGACCACAGTGCT 253bp chr17:73840245-73840497
2-3-4 TCAGAGGAGCCATTCCTGTC CCAGAATGCCCTTGGCCTGT 723bp chr17:73838953-73839675
5-6 CAGACACTGCAGCGGGTCA CCTGAGCCAGGACGACCT 708bp chr17:73838417-73839124
7-8-9 TCTGGGCTGTGGTCACTTAC GACAGGAGGTGGACCCCA chr17:73837143-?
10-11-12 GGGTCCACCTCCTGTCG TACACTTTGGGCTACCGGC chr17:?-73835853
13-14-15 TGGATGGGAGGCGATTC GGTGGAACCATTCAGTGGTGC 782bp chr17:73832313-73833094
16-17-18 CCATTGCCCCAGCTGGTGA TGTGGTCCTGCACCCGCTT 619bp chr17:73831834-73832452
19-20 CCTCTTCTCCATGGCTTTCCG GGGATGTTCAGAGCGGGTTAC 593bp chr17:73831380-73831972
21-22 TCTGTGCCTGGTGATGGTAG GCTTCCTGAACTGTGCCC 607bp chr17:73830631-73831237
23-24 CCAACATGGTAAGGCCAGAG ACCAAGGCCTTCAATGACC 626bp chr17:73830081-73830706
25 CTCCAGGTCAGGGGAAGG CCTCAGAACGGATGCTGG 248bp chr17:73828956-73829203
26-27 AGAGCGTCTTTGCTTCCTCC TTATGTAGCCGACCCTGGAC 407bp chr17:73827094-73827500
28-29 AAACGGACATGTTACCCTTGAC CTCCTGCAGGGTGAGCGAA 491bp chr17:73826352-73826842
30 AGCATAGCTGCTGGGGTG CATCTCAGGGGAGCCCCTT 296bp chr17:73826005-73826300
31 CCAACACCAAGTGGGTGAAC GACCCTGGCATCACCTCTC 365bp chr17:73824783-73825147
32 GGAGCAGTGTTGTGTACCCC TCTGGGCACTCCGCATGCT 359bp chr17:73823906-73824264

IV. Ethanol precipitation

V. Sequencing analysis