PCR设计引物时酶切位点的保护碱基表

切割率% 酶 寡核苷酸序列 2 hr Acc I 20 hr GGTCGACC CGGTCGACCG CCGGTCGACCGG 0 0 0 0 >90 >90 >90 >90 >90 50 >90 >90 10 >90 >90 0 75 25 0 0 50 0 0 25 25 0 0 >90 50 >90 >90 >90 0 0 0 0 >90 >90 >90 >90 >90 >90 >90 >90 25 >90 >90 0 >90 >90 0 0 >90 10 0 50 >90 0 0 >90 50 >90 >90 >90 Afl III CACATGTG CCACATGTGG CCCACATGTGGG Asc I GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA Ava I CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA BamH I CGGATCCG CGGGATCCCG CGCGGATCCGCG Bgl II CAGATCTG GAAGATCTTC GGAAGATCTTCC BssH II GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA BstE II BstX I GGGT(A/T)ACCC AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT Cla I CATCGATG GATCGATC CCATCGATGG CCCATCGATGGG EcoR I GGAATTCC CGGAATTCCG CCGGAATTCCGG Hae III GGGGCCCC AGCGGCCGCT TTGCGGCCGCAA >90 >90 >90 0 0 10 0 >90 >90 0 25 0 50 0 0 0 0 75 75 0 10 10 >90 >90 >90 0 0 75 0 >90 >90 0 50 0 75 0 0 0 0 >90 >90 0 25 50 Hind III CAAGCTTG CCAAGCTTGG CCCAAGCTTGGG Kpn I GGGTACCC GGGGTACCCC CGGGGTACCCCG Mlu I GACGCGTC CGACGCGTCG Nco I CCCATGGG CATGCCATGGCATG Nde I CCATATGG CCCATATGGG CGCCATATGGCG GGGTTTCATATGAAACCC GGAATTCCATATGGAATTCC GGGAATTCCATATGGAATTCCC Nhe I GGCTAGCC CGGCTAGCCG CTAGCTAGCTAG PCR设计引物时酶切位点的保护碱基表2

切割率% 酶 寡核苷酸序列 2 hr Not I 20 hr TTGCGGCCGCAA ATTTGCGGCCGCTTTA AAATATGCGGCCGCTATAAA ATAAGAATGCGGCCGCTAAACTAT AAGGAAAAAAGCGGCCGCAAAAGGAAAA 0 10 10 25 25 10 >90 0 0 0 0 10 10 90 >90 >90 >90 0 25 >90 Nsi I TGCATGCATGCA CCAATGCATTGGTTCTGCAGTT Pac I TTAATTAA GTTAATTAAC CCTTAATTAAGG Pme I GTTTAAAC GGTTTAAACC GGGTTTAAACCC AGCTTTGTTTAAACGGCGCGCCGG 0 0 0 75 0 10 >90 >90 0 0 10 0 10 0 50 0 10 10 0 25 50 >90 0 10 >90 >90 0 0 25 10 10 0 >90 0 50 75 Pst I GCTGCAGC TGCACTGCAGTGCA AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT Pvu I CCGATCGG ATCGATCGAT TCGCGATCGCGA Sac I Sac II CGAGCTCG GCCGCGGC TCCCCGCGGGGA Sal I GTCGACGTCAAAAGGCCATAGCGGCCGC GCGTCGACGTCTTGGCCATAGCGGCCGCGG ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA Sca I GAGTACTC AAAAGTACTTTT 10 75 0 0 10 >90 10 10 0 0 0 0 10 >90 >90 >90 25 75 10 10 50 >90 >90 >90 50 50 0 25 50 >90 >90 >90 Sma I CCCGGG CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA Spe I GACTAGTC GGACTAGTCC CGGACTAGTCCG CTAGACTAGTCTAG Sph I GGCATGCC CATGCATGCATG ACATGCATGCATGT Stu I AAGGCCTT GAAGGCCTTC AAAAGGCCTTTT Xba I CTCTAGAG GCTCTAGAGC TGCTCTAGAGCA CTAGTCTAGACTAG 0 >90 75 75 0 10 10 0 25 50 >90 0 >90 >90 >90 0 25 75 0 75 >90 >90 Xho I CCTCGAGG CCCTCGAGGG CCGCTCGAGCGG Xma I CCCCGGGG CCCCCGGGGG CCCCCCGGGGGG TCCCCCCGGGGGGA