Table 3 of
Dekomien, Mol Vis 2002;
8:138-142.
Table 3. Primers and conditions for PCR amplification
PCR systems used to characterize the canine PDC gene including mutation screening. Systems 1aUTRF, 2(1b)2F, 3(2)R, 3(2)F, 4(3)1R, 4(3)F, 4(3)1R, 4(3)2F, and 4(3)2R use PCR primer sequences selected from the canine mRNA (EMBL accession number Y17697) for the identification of exon/intron boundaries and intron sequences. Systems 2(1b)2F, 2(1b)IR, 3(1b)IF, 3(2) IR, 4(3)1IF, 4(3)1R, 4(3)2F, and 4(3)2R use PCR primers selected on the basis of the canine PDC gene (EMBL accessions numbers CFA417559 through CFA17561) for mutation analysis.
Fragment Annealing T/MgCl2 PCR system Primer sequence (5'->3') Location length (bp) [(°C)/(mM)] ---------- ------------------------ --------- ----------- ----------------- 1a UTR F AATTCGGCACGAAACAAGC E1 ~14000 59/1.5 2 (1) R CTGTATGTGTGGCCTGTC E2 2 (1a)IF ACTCCTACCCTAGCATGCA I1 191 54/1 2 (1) R CTGTATGTGTGGCCTGTC E2 2 (1b)2F AGATTATATAAAATCAAATCC E2 169 51/1 2 (1b)IR TTTATGCTGACCAATGGCAGT I2 2 (1b)2F AGATTATATAAAATCAAATCC E2 ~1900 54/2* 3 (2) R ACCTTTCTGCTGAATCTTTCTT E3 3 (2) F AGGACCCAAAGGAGTAATAAA E3 ~1500 54/2* 4 (3)1R CCCACCTTTGTAGATGAGCA E4 3 (1b)IF TCTGATGCAGAAGTTGATATCT I2 396 56/1 3 (2)IR TTTGCCAATGTAACCAATACTAT I3 4 (3) F AGATGAGCATTCAAGAATATGA E4 374 55/1 4 (3)1R CCCACCTTTGTAGATGAGCA E4 4 (3)1IF AAAGCCTGTTTCTCTATCT I3 509 53/1** 4 (3)1R CCCACCTTTGTAGATGAGCA E4 4 (3)2F GCTGGGGACCGCTTTTCCTCAGA E4 512 56/1 4 (3)2R AAACAAGGAGTAAAAAAAGTCAAC 3' UTR E4 *Also contains 5% DMSO **Also contains 5% Formamide |