Table 1 of
Otteson, Mol Vis 2005;
11:986-995.
Table 1. Primers for PCR amplification of zebrafish crx from genomic DNA
Oligonucleotide primers used for PCR amplification of zebrafish crx for sequence analysis were designed using Primer3 (Primer3) or Vector NTI (Invitrogen Corp.) software and the published zebrafish crx cDNA sequence (GenBank accession number AF503443) [4] and obtained from IDT as standard, desalted oligonucleotides (25 nmole synthesis). Annealing temperatures were optimized using a gradient program on a MJ Research DNA Engine thermocycler (BioRad).
Temp (anneal) Size Forward primer (5'-3') Reverse primer (5'-3') Target (°C) (bp) ------------------------- --------------------------- ---------- -------- --------- CTCATAGTGTCCCGCTCCTGTCC GCTCTCTCTCTCCCTCCCTCTCTCAC exon 1:5' 60 364 TTCTCGCATATAGACATTTATTTG ATAAAACGTCTAACAGAAACACCATTA exon 1:3' 55 471 AACCTCCTGAATTGTTTTAAGCTG TGTTGAGAAGGATGTGTGAGAGGC exon 2 55 242 ATCCATCCATTCATCCATCTAACAC CTCTCTCACTTTCTTTTCTACAGCAC exon 3:5' 55 990 ACGGTCAGCCCTCTTCCTACAGCC GGAAGGCTCAGATATAGGGGTGG exon 3:mid 60 725 GGAAGGAAAGATGCGTGAAGAAG AACCTTCAGCTCCCCTCTGTACGTG exon 3:3' 60 933 TTCTCGCATATAGACATTTATTTG TGTCTGGATAGCGTGTTTTGGTGA intron 1 62 1243 ACCACCTTCACTCGCACCCAG TGTAGGAAGAGGGCTGACCGTAGC intron 2 62 1026-1064 AACCTGCCAGAGGGTTACT TCAACAGATGGCTTCAGTGC Z13833 50 177 [33] |