酶切位点保护碱基.doc

。PCR设计引物时酶切位点的保护酶寡核苷酸序列切割率%2 hr20 hrAcc IGGTCGACC CGGTCGACCG CCGGTCGACCGG 0 0 00 0 0Afl IIICACATGTG CCACATGTGG CCCACATGTGGG0 90 900 90 90Asc IGGCGCGCC AGGCGCGCCT TTGGCGCGCCAA90 90 9090 90 90Ava ICCCCGGGG CCCCCGGGGG TCCCCCGGGGGA50 90 9090 90 90BamH ICGGATCCG CGGGATCCCG CGCGGATCCGCG10 90 9025 90 90Bgl IICAGATCTG GAAGATCTTC GGAAGATCTTCC0 75 250 90 90BssH IIGGCGCGCC AGGCGCGCCT TTGGCGCGCCAA0 0 500 0 90BstE IIGGGT(A/T)ACCC010BstX IAACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT0 25 250 50 90Cla ICATCGATG GATCGATC CCATCGATGG CCCATCGATGGG0 0 90 500 0 90 50EcoR IGGAATTCC CGGAATTCCG CCGGAATTCCGG90 90 9090 90 90Hae IIIGGGGCCCC AGCGGCCGCT TTGCGGCCGCAA90 90 9090 90 90Hind IIICAAGCTTG CCAAGCTTGG CCCAAGCTTGGG0 0 100 0 75Kpn IGGGTACCC GGGGTACCCC CGGGGTACCCCG0 90 900 90 90Mlu IGACGCGTC CGACGCGTCG0 250 50Nco ICCCATGGG CATGCCATGGCATG0 500 75Nde ICCATATGG CCCATATGGG CGCCATATGGCG GGGTTTCATATGAAACCC GGAATTCCATATGGAATTCC GGGAATTCCATATGGAATTCCC0 0 0 0 75 750 0 0 0 90 90Nhe IGGCTAGCC CGGCTAGCCG CTAGCTAGCTAG0 10 100 25 50Not 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 ICCTCGAGG CCCTCGAGGG CCGCTCGAGCGG0 10 100 25 75Xma ICCCCGGGG CCCCCGGGGG CCCCCCGGGGGG TCCCCCCGGGGGGA0 25 50 900 75 90 90注释：1如果要加在序列的5端，就在酶切位点识别碱基序列（红色）的5端加上相应的碱基（黑色），相同如果要在3端加保护碱基，就在酶切位点