PCR设计引物时酶切位点的保护碱基表1

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%2hr20hrAccIGGTCGACCCGGTCGACCGCCGGTCGACCGG000000AflIIICACATGTGCCACATGTGGCCCACATGTGGG0909009090AscIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA909090909090AvaICCCCGGGGCCCCCGGGGGTCCCCCGGGGGA509090909090PCR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%2hr20hrBamHICGGATCCGCGGGATCCCGCGCGGATCCGCG109090259090BglIICAGATCTGGAAGATCTTCGGAAGATCTTCC0752509090BssHIIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA00500090BstEIIGGGT(A/T)ACCC010BstXIAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCATTGGATGCAT0252505090PCR设计引物时酶切位点的保护碱基表3酶寡核苷酸序列切割率%2hr20hrClaICATCGATGGATCGATCCCATCGATGGCCCATCGATGGG009050009050EcoRIGGAATTCCCGGAATTCCGCCGGAATTCCGG909090909090HaeIIIGGGGCCCCAGCGGCCGCTTTGCGGCCGCAA909090909090HindIIICAAGCTTGCCAAGCTTGGCCCAAGCTTGGG00100075KpnIGGGTACCCGGGGTACCCCCGGGGTACCCCG0909009090PCR设计引物时酶切位点的保护碱基表4酶寡核苷酸序列切割率%2hr20hrMluIGACGCGTCCGACGCGTCG025050NcoICCCATGGGCATGCCATGGCATG050075NdeICCATATGGCCCATATGGGCGCCATATGGCGGGGTTTCATATGAAACCCGGAATTCCATATGGAATTCCGGGAATTCCATATGGAATTCCC0000757500009090NheIGGCTAGCCCGGCTAGCCGCTAGCTAGCTAG0101002550PCR设计引物时酶切位点的保护碱基表5酶寡核苷酸序列切割率%2hr20hrNotITTGCGGCCGCAAATTTGCGGCCGCTTTAAAATATGCGGCCGCTATAAAATAAGAATGCGGCCGCTAAACTATAAGGAAAAAAGCGGCCGCAAAAGGAAAA010102525010109090NsiITGCATGCATGCACCAATGCATTGGTTCTGCAGTT10909090PacITTAATTAAGTTAATTAACCCTTAATTAAGG00002590PmeIGTTTAAACGGTTTAAACCGGGTTTAAACCCAGCTTTGTTTAAACGGCGCGCCGG000750255090PstIGCTGCAGCTGCACTGCAGTGCAAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCATTGGATGCAT0109090001090900酶切位点保护碱基表6酶寡核苷酸序列切割率%2hr20hrPvuICCGATCGGATCGATCGATTCGCGATCGCGA010002510SacICGAGCTCG1010SacIIGCCGCGGCTCCCCGCGGGGA050090SalIGTCGACGTCAAAAGGCCATAGCGGCCGCGCGTCGACGTCTTGGCCATAGCGGCCGCGGACGCGTCGACGTCGGCCATAGCGGCCGCGGAA0101005075ScaIGAGTACTCAAAAGTACTTTT10752575SmaICCCGGGCCCCGGGGCCCCCGGGGGTCCCCCGGGGGA00109010105090SpeIGACTAGTCGGACTAGTCCCGGACTAGTCCGCTAGACTAGTCTAG10100090905050酶切位点保护碱基表7酶寡核苷酸序列切割率%2hr20hrSphIGGCATGCCCATGCATGCATGACATGCATGCATGT001002550StuIAAGGCCTTGAAGGCCTTCAAAAGGCCTTTT909090909090XbaICTCTAGAGGCTCTAGAGCTGCTCTAGAGCACTAGTCTAGACTAG09075750909090XhoICCTCGAGGCCCTCGAGGGCCGCTCGAGCGG0101002575XmaICCCCGGGGCCCCCGGGGGCCCCCCGGGGGGTCCCCCCGGGGGGA02550900759090由于直接暴露在末端的酶切位点不容易直接被限制性核酸内切酶切开，因此在设计PCR引物时，人为的在酶切位点序列的5‘端外侧添加额外的碱基序列，即保护碱基，用来提高将来酶切时的活性。