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

Dekomien, Mol Vis 2002; 8:138-142 <http://www.molvis.org/molvis/v8/a19/>
©2002 Molecular Vision <http://www.molvis.org/molvis/>
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