Table 2 of
Dekomien, Mol Vis 2002;
8:436-441.Table 2 of
Dekomien, Mol Vis 2002;
8:436-441.
Table 2. Primers and conditions for PCR amplification
PCR systems to characterize the canine RCV1 gene including mutation screening. Systems designated by H: PCR primer sequences selected from the human mRNA (EMBL accession number AB001838) for the identification of exon/intron boundaries and canine gene sequences. Systems designated by I: PCR primers (on the basis of the canine RCV1 gene EMBL accession number AJ414401) to amplify the complete intron. Systems designated by M: PCR primers (on the basis of the canine RCV1 gene) for mutation analysis. Ht: Additional PCR primers used for haplotype analysis. *Nucleotide position to which the 5' end of the primer (red type) hybridizes. ** Addition of 5% DMSO.
Restriction Annealing
PCR Fragment enzymes for T/MgCl2
PCR-system Primer sequence (5'->3')* Location length (bp) SSCP analysis (°C/mM)
---------- ------------------------- ----------- ------------------ ------------- ---------
H 1F CCCTGTCCAAGGAGATCCT E 1; 13 359 - 47/2**
H 1R CATGACGATCTCCAGCACTT E 1; 381
I 1-331EF TCTCGCTCTACGACGTGGA E 1; 580 837 - 54/2
I 1-IR GACAGGGAACGGAGTGAAG I 1; 1419
H 1F CCCTGTCCAAGGAGATCCT E 1; 13 1422 - 46/2**
H 2R CTTTCCAAAGTACTTCCAGATC E 2; 483
H 2F TTCAAAATGATCACTCCCGAGG E 2; 388 2684 - 48/2**
H 3R TCATCTTTTCCTTCACTTTTTGAG E 3; 592
I 2F ACCGCCGAGCTCCCTGG I 2; 2006 1774 - 54/2**
I 2R ATGCGGAGACACTCCCACG I 2; 3777
I/M 2 2F TGAGAACTGGCCTTCGAGG I 2; 2398 1020: 208, 98, 160, DdeI 52/1
I/M 2 2R CGTGACTGTGGCATTCACC I 2; 3417 13, 188, 276, 77
I/M 2Ht 2F GGGAGGCATTCCAGAACAG I 2; 2658 760: 30, 160, 13, DdeI 54/1
I/M 2 2R CGTGACTGTGGCATTCACC I 2; 3417 188, 276, 77
M 5'-UTR1F CTCCCTGAAGGCCAAGATG UTR/E 1; 1 372: 184, 132, PstI 55/1
M/Ht Kl1ER TTCGGAAGGAAACTGGTACC E 1; 374 56
H/M 1F CCCTGTCCAAGGAGATCCT E 1; 13 424: 183, 71, NlaIII 54/1
M 1-IR GAACCCCCTGGACCCAGGA I 1; 709 145, 65
M 2-IF GGAATCTGTATTTTCACCTCTG I 1; 1483 308: 136, DdeI 55/1
M 2-IR CTAGGAAGACCAATCTCACT I 2; 1790 172
M 3-IF GTGATGGGTTGTACCCTCAG I 2; 4019 340: 206, MboI 55/1
M UTR 3'R GCATGTGCACGTGCTCACG 3'-UTR; 4358 134
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