AIRE
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 TTTGCTCTTTGCGTGGTCG ACTTGTCATCAGGGACGCC 478bp chr21:45705704+45706181
2 CACCTGACTCCACCACAAGC AAGACGGTCCTGGAACATCAC 454bp chr21:45706285+45706738
3-4 ACCTGTCTGGCCAAGGTGTC AGACCCAAGGTGTCCCCTTC 885bp chr21:45706769+45707653
5 TTCCTATCCATTGTGCCAGC CAAGGACGCTATGCTACAGG 760bp chr21:45707966+45708725
6-7 TGTTGAACACCCTGGTGTAG GACCTCACTTTCTCTGACTC 814bp chr21:45709349+45710162
8a CACCAGGTAATGCCCTAGAC CCACATGAACCCTTCCTCAC 750bp chr21:45709946+45710695
8b AATGAGTGAGGTCATTGCCG TTCTGCTTTGCTGCCCTCAG 859bp chr21:45710476+45711334
9 GGATTCCTGGGGTTCATCAG GCACAGTAGACCTACTACCC 670bp chr21:45711936+45712605
10 GGACCACAGAGCCAGGAAG GCTGGAGTGGAGGTCTAGGAG 572bp chr21:45712677+45713248
11 CCTCACTTGCGCCTAGACC TGGGCTGTATGATGTGGGTG 338bp chr21:45713544+45713881
12 ATACCCCGGAGGTGGCACT AGGACAGGAAGAGCTACTGG 643bp chr21:45714221+45714863
13 ACACAGTGTGGAGGCTGTCTG AGGTGCTCAGCAGCACTGG 474bp chr21:45716026+45716499
14 CGATGGCCATGATTCTGTG ACTTCCATGAGGTCCCTGG 783bp chr21:45717470+45718252

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 TTTGCTCTTTGCGTGGTCG ACTTGTCATCAGGGACGCC 478bp chr21:45705704+45706181
2 CACCTGACTCCACCACAAGC AAGACGGTCCTGGAACATCAC 454bp chr21:45706285+45706738
3-4 ACCTGTCTGGCCAAGGTGTC AGACCCAAGGTGTCCCCTTC 885bp chr21:45706769+45707653
5 TTCCTATCCATTGTGCCAGC CAAGGACGCTATGCTACAGG 760bp chr21:45707966+45708725
6-7 TGTTGAACACCCTGGTGTAG GACCTCACTTTCTCTGACTC 814bp chr21:45709349+45710162
8a CACCAGGTAATGCCCTAGAC CCACATGAACCCTTCCTCAC 750bp chr21:45709946+45710695
8b AATGAGTGAGGTCATTGCCG TTCTGCTTTGCTGCCCTCAG 859bp chr21:45710476+45711334
9 GGATTCCTGGGGTTCATCAG GCACAGTAGACCTACTACCC 670bp chr21:45711936+45712605
10 GGACCACAGAGCCAGGAAG GCTGGAGTGGAGGTCTAGGAG 572bp chr21:45712677+45713248
11 CCTCACTTGCGCCTAGACC TGGGCTGTATGATGTGGGTG 338bp chr21:45713544+45713881
12 ATACCCCGGAGGTGGCACT AGGACAGGAAGAGCTACTGG 643bp chr21:45714221+45714863
13 ACACAGTGTGGAGGCTGTCTG AGGTGCTCAGCAGCACTGG 474bp chr21:45716026+45716499
14 CGATGGCCATGATTCTGTG ACTTCCATGAGGTCCCTGG 783bp chr21:45717470+45718252

IV. Ethanol precipitation

V. Sequencing analysis