保护碱基表.doc

不同内切酶对识别位点以外最少保护碱基数目的要求 PCR设计引物时酶切位点的保护碱基表1 酶 寡核苷酸序列 切割率% 2 hr 20 hr Acc I GGTCGACC CGGTCGACCG CCGGTCGACCGG 0 0 0 0 0 0 Afl III CACATGTG CCACATGTGG CCCACATGTGGG 0 90 90 0 90 90 Asc I GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA 90 90 90 90 90 90 Ava I CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA 50 90 90 90 90 90 BamH I CGGATCCG CGGGATCCCG CGCGGATCCGCG 10 90 90 25 90 90 Bgl II CAGATCTG GAAGATCTTC GGAAGATCTTCC 0 75 25 0 90 90 BssH II GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA 0 0 50 0 0 90 BstE II GGGT(A/T)ACCC 0 10 BstX I AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT 0 25 25 0 50 90 Cla I CATCGATG GATCGATC CCATCGATGG CCCATCGATGGG 0 0 90 50 0 0 90 50 EcoR I GGAATTCC CGGAATTCCG CCGGAATTCCGG 90 90 90 90 90 90 Hae III GGGGCCCC AGCGGCCGCT TTGCGGCCGCAA 90 90 90 90 90 90 Hind III CAAGCTTG CCAAGCTTGG CCCAAGCTTGGG 0 0 10 0 0 75 Kpn I GGGTACCC GGGGTACCCC CGGGGTACCCCG 0 90 90 0 90 90 Mlu I GACGCGTC CGACGCGTCG 0 25 0 50 Nco I CCCATGGG CATGCCATGGCATG 0 50 0 75 Nde I CCATATGG CCCATATGGG CGCCATATGGCG GGGTTTCATATGAAACCC GGAATTCCATATGGAATTCC GGGAATTCCATATGGAATTCCC 0 0 0 0 75 75 0 0 0 0 90 90 Nhe I GGCTAGCC CGGCTAGCCG CTAGCTAGCTAG 0 10 10 0 25 50 PCR设计引物时酶切位点的保护碱基表2酶寡核苷酸序列切割率%2 hr20 hrNot ITTGCGGCCGCAA ATTTGCGGCCGCTTTA AAATATGCGGCCGCTATAAA ATAAGAATGCGGCCGCTAAACTAT AAGGAAAAAAGCGGCCGCAAAAGGAAAA0 10 10 25 250 10 10 90 90Nsi ITGCATGCATGCA CCAATGCATTGGTTCTGCAGTT10 9090 90Pac ITTAATTAA GTTAATTAAC CCTTAATTAAGG0 0 00 25 90Pme IGTTTAAAC GGTTTAAACC GGGTTTAAACCC AGCTTTGTTTAAACGGCGCGCCGG0 0 0 750 25 50 90Pst IGCTGCAGC TGCACTGCAGTGCA AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT0 10 90 90 00 10 90 90 0Pvu ICCGATCGG ATCGATCGAT TCGCGATCGCGA0 10 00 25 10Sac ICGAGCTCG1010Sac IIGCCGCGGC TCCCCGCGGGGA0 500 90Sal IGTCGACGTCAAAAGGCCATAGCGGCCGC GCGTCGACGTCTTGGCCATAGCGGCCGCGG ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA0 10 100 50 75Sca IGAGTACTC AAAAGTACTTTT10 7525 75Sma ICCCGGG CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA0 0 10 9010 10 50 90Spe IGACTAGTC GGACTAGTCC CGGACTAGTCCG CTAGACTAGTCTAG10 10 0 090 90 50 50Sph IGGCATGCC CATGCATGCATG ACATGCATGCATGT0 0 100 25 50Stu IAAGGCCTT GAAGGCCTTC AAAAGGCCTTTT90 90 9090 90 90Xba ICTCTAGAG GCTCTAGAGC TGCTCTAGAGCA CTAGTCTAGACTAG0 90 75 750 90 90 90Xho ICCT