Table 3 of
Dekomien, Mol Vis 2003;
9:601-605.Table 3 of
Dekomien, Mol Vis 2003;
9:601-605.
Table 3. Primers and conditions for PCR amplification
PCR protocols used to characterize the canine RPE65 gene including mutation screening. For RPE65 sequences, see EMBL accession entries AJ506753, AJ506754, AJ506755, AJ506756, AJ506757, AJ506758, AJ506759, and AJ251207.
Annealing
Fragment T/MgCl2/ Restriction
length formamide enzymes for
PCR-system Location Primer sequenceA (5'->3') (bp) [(°C)/(mM)/%] SSCP-analyses
------------- --------- ------------------------- -------- ------------- -------------
RPE65UTR 1-F Exon 1 CGACCGTCTGTCCTGCCC 186 56/1.5/5 -
RPE65 I-1-R Intron 1 GGGTATTTGCTTCAATCCATG
RPE65UTR21F Exon 1 CTGGGAGACAATGTCCATC 1124 51/1.5 -
RPE65 E2-2R Exon 2 CGACGACAGCTCTTCCAC
RPE65 E4 F Exon 4 AGGTTCATCCGCACCGATG 357 54/1.5 AluI/HincII
RPE65 E5 R Exon 5 ACCTGCTTAATTGTCTCCAG
RPE65 I 4 F Intron 4 CACAGCTTGAAGGTTACTGGAC 286 54/1.5 HincII
RPE65 I 5 R Intron 5 TCTATTTGGCCCTCATGAGC
RPE65 E5 F Exon 5 AGGTTTTTTTCTTACTTCCGA ~2800 60/1.5 -
RPE65 E6 R Exon 6 TGCTTGGAGTGGAGGGATC
RPE65 E-6-F Exon 6 AGGTTGATCTCTGCAACTAC 377 56/1.5 DdeI
RPE65 I 6 R Intron 6 CTGTGGAATACGACTTGGC
RPE65 E-6-F Exon 6 AGGTTGATCTCTGCAACTAC ~2000 60/1.0 -
RPE65 E-7-R Exon 7 CTATGGACGTACGATGGC
RPE65 7EF Exon 7 AGACAAGGAAGATCCAATAAGC 443 54/1.5 TruI
RPE65 8ER Exon 8 CCCATGGTTTCATTGGACTC
RPE65 8EF Exon 8 TTTTGTGGAGACGCCAGTC 369 60/2.5 BsiHKAI
RPE65 9ER Exon 9 CCTTTCCAGCAGCAGAGATC
RPE65 E-9-F Exon 9 AGGTTTGGCTTCACATCGCTG ~2000 60/1.0 -
RPE65 E-10-R Exon 10 TTGTCGATATTCAGAGGAAGC
RPE65 I-9 F Intron 9 ACACCCCAGAGCAAGCAGG 229 59/1.5 EcoRI
RPE65 E-10-R Exon 10 TTGTCGATATTCAGAGGAAGC
RPE65 E11-F Exon 11 GCCGACACAGGCAAGAAC 542 55/1.5 DdeI
RPE65 E12-R Exon 12 ACCCTGTCCGGAACGAAG
RPE65 E-12-F Exon 12 CCTCAAATCAACTATCAGAAGTC 580 58/1.0 -
RPE65 I-13-R Intron 13 AACACACTAACATAGAGAACTC
RPE65 I-12 F Intron 12 AAGAGAAAAGTAGTTTGAGTCAC 261 57/1.5 HphI
RPE65 I-13-R Intron 13 AACACACTAACATAGAGAACTC
RPE65 E13-F Exon 13 CTCTGCAAGCTGAACGTC ~1800 55/1.0 -
RPE65 E/I14R Exon 14 CCATGTAATACACAGCAGGCTAA
RPE65 E 14-F Exon 14 AGGTGTAGTTCTGAGTGTG 249 55/1.0 RsaI
RPE65 E 14-R Exon 14 CCGTATACAGCAGGCTAAA
RPE65 UTR14F Exon 14 CAAAGTCAAGAAAAAGTGAGGT 705 53/2.0 MseI
RPE65 UTR14R Exon 14 GCTTTGATGTTATGTAAGCTTTT
RPE65 UTR14F Exon 14 CAAAGTCAAGAAAAAGTGAGGT 277 55/1.5 MnlI
RPE65 UTR214R Exon 14 CACTTTCATAATAGGAACAAGAA
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