Table 2

List of primers

Primer nameSequenceDerived from GenBank accession number
To allow the discrimination between alleles D114 and 114Y (340G→T) the primers pCANmF and pCANmR were modified in the following way. For pCANmF the nine nucleotides at the 5` end are derived from the 3` splice acceptor of intron 1.4 In order to create an internal SnaBI control restriction site nucleotides C21, A22, and A24 (in bold, numbered from the 5` end) were replaced with A, C, and T (underlined), respectively. For pCANmR, nucleotides A25 and G27 were replaced with T and C (underlined), respectively. These replacements allowed SnaBI digestion of the mutant 114Y allele while leaving the D114 allele resistant.
cDNA primers
CAF15`CTGTACTTTGCCCTTTTGGTAS67156.1
CAF35`CTGATTGAGCATCCTTCCCTCAS67156.1
CAR35`ACACCGTGTAAGATGTAAGCTS67156.1
Genomic DNA primers
CAex1F5`CTTCTGAATTGCAGAAATCAGAI70466
CAex1R5`CACACCTACCACTTTCACACAA
CAex2F5`CTCAGGCACAGATGTTGTTCAI70467
CAex2R5`CAAGTCCTTTGCTGACTTATAA
CAex3F5`GGGTTTTTACCCAAGAAAGACGTI70468
CAex3R5`CTCTGAGTTTCAGCTAGGACAT
CAex4F5`ATGTGACTATCTCCTTCTGTI70469
CAex4R5`TCTGACCCAGGTTCCAATTGTTA
CAex5F5`CCAGAGATGTTTTTAGTTGCCAI70470
CAex5R5`TGCTGTATGAGCTATAAACTTCT
CAex6F5`GAGTCTGACATAAATTTTTAGAGGAI70471
pCANmF5`TTATAACAGCAAAAAAATGTACGTAGATTTGCCATATGAAGTG
pCANmR5`CCCCATGTTAGAGGTGGTGTTGTGTACGT