PCR设计引物时酶切位点的保护碱基表

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1、PCR设计引物时酶切位点的保护碱基表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%2hr20hrAccIGGTCGACCCGGTCGACCGCCGGTCGACCGG000000AflIIICACATGTGCCACATGTGGCCCACATGTGGG0>90>900>90>90AscIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA>90>90>90>90>90>90AvaICCCCGGGGCCCCCGGGGGTCCCCCGGGG

2、GA50>90>90>90>90>90BamHICGGATCCGCGGGATCCCGCGCGGATCCGCG10>90>9025>90>90BglIICAGATCTGGAAGATCTTCGGAAGATCTTCC075250>90>90BssHIIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA005000>90BstEIIGGGT(A/T)ACCC010BstXIAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCAT

3、TGGATGCAT02525050>90ClaICATCGATGGATCGATCCCATCGATGGCCCATCGATGGG00>905000>9050EcoRIGGAATTCCCGGAATTCCGCCGGAATTCCGG>90>90>90>90>90>90HaeIIIGGGGCCCCAGCGGCCGCTTTGCGGCCGCAA>90>90>90>90>90>90HindIIICAAGCTTGCCAAGCTTGGCCCAAGCTTGGG00100075KpnIGGGTACCCGGGGTACCCCCG

4、GGGTACCCCG0>90>900>90>90MluIGACGCGTCCGACGCGTCG025050NcoICCCATGGGCATGCCATGGCATG050075NdeICCATATGGCCCATATGGGCGCCATATGGCGGGGTTTCATATGAAACCCGGAATTCCATATGGAATTCCGGGAATTCCATATGGAATTCCC000075750000>90>90NheIGGCTAGCCCGGCTAGCCGCTAGCTAGCTAG0101002550PCR设计引物时酶切位点

5、的保护碱基表2酶寡核苷酸序列切割率%2hr20hrNotITTGCGGCCGCAAATTTGCGGCCGCTTTAAAATATGCGGCCGCTATAAAATAAGAATGCGGCCGCTAAACTATAAGGAAAAAAGCGGCCGCAAAAGGAAAA0101025250101090>90NsiITGCATGCATGCACCAATGCATTGGTTCTGCAGTT10>90>90>90PacITTAATTAAGTTAATTAACCCTTAATTAAGG000025>90PmeIGTTTAAAC

6、GGTTTAAACCGGGTTTAAACCCAGCTTTGTTTAAACGGCGCGCCGG0007502550>90PstIGCTGCAGCTGCACTGCAGTGCAAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCATTGGATGCAT010>90>900010>90>900PvuICCGATCGGATCGATCGATTCGCGATCGCGA010002510SacICGAGCTCG1010SacIIGCCGCGGCTCC

7、CCGCGGGGA0500>90SalIGTCGACGTCAAAAGGCCATAGCGGCCGCGCGTCGACGTCTTGGCCATAGCGGCCGCGGACGCGTCGACGTCGGCCATAGCGGCCGCGGAA0101005075ScaIGAGTACTCAAAAGTACTTTT10752575SmaICCCGGGCCCCGGGGCCCCCGGGGGTCCCCCGGGGGA0010>90101050>90SpeIGACTAGTCGGACTAGTCCCGGACTAGTCCGCTAGACTAGT

8、CTAG101000>90>905050SphIGGCATGCCCATGCATGCATGACATGCATGCATGT001002550StuIAAGGCCTTGAAGGCCTTCAAAAGGCCTTTT>90>90>90>90>90>90XbaICTCTAGAGGCTCTAGAGCTGCTCTAGAGCACTAGTCTAGACTAG0>9075750>90>90>90XhoICCTCGAGGCCCTCGAGGGCCGCTCGAGCGG0101002575XmaICCC