PCR设计引物时酶切位点的保护.doc

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1、PCR设计引物时酶切位点的保护酶寡核苷酸序列切割率%2hr20hrAccIGGTCGACCCGGTCGACCGCCGGTCGACCGG000000AflIIICACATGTGCCACATGTGGCCCACATGTGGG0>90>900>90>90AscIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA>90>90>90>90>90>90AvaICCCCGGGGCCCCCGGGGGTCCCCCGGGGGA50>90>90>90>90>90BamHICGGATCCGCGGGATCCCGCGCGGATCCGCG10>90>9025>90>90BglIICAGATCTG

2、GAAGATCTTCGGAAGATCTTCC075250>90>90BssHIIGGCGCGCCAGGCGCGCCTTTGGCGCGCCAA005000>90BstEIIGGGT(A/T)ACCC010BstXIAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCATTGGATGCAT02525050>90ClaICATCGATGGATCGATCCCATCGATGGCCCATCGATGGG00>905000>9050EcoRIGGAATTCCCGGAATTCCGCCGGAATTCCGG>90>90>90>

3、90>90>90HaeIIIGGGGCCCCAGCGGCCGCTTTGCGGCCGCAA>90>90>90>90>90>90HindIIICAAGCTTGCCAAGCTTGGCCCAAGCTTGGG00100075KpnIGGGTACCCGGGGTACCCCCGGGGTACCCCG0>90>900>90>90MluIGACGCGTCCGACGCGTCG025050NcoICCCATGGGCATGCCATGGCATG050075NdeICCATATGGCCCATATGGGCGCCATATGGCGGGGTTTCATATGAAACCCGGAATTCCATATGGAATTCCGGG

4、AATTCCATATGGAATTCCC000075750000>90>90NheIGGCTAGCCCGGCTAGCCGCTAGCTAGCTAG0101002550NotITTGCGGCCGCAAATTTGCGGCCGCTTTAAAATATGCGGCCGCTATAAAATAAGAATGCGGCCGCTAAACTATAAGGAAAAAAGCGGCCGCAAAAGGAAAA0101025250101090>90NsiITGCATGCATGCACCAATGCATTGGTTCTGCAGTT10>90>90>90PacITTAATTAAGTTAATTAACCCTTAATTAAGG000

5、025>90PmeIGTTTAAACGGTTTAAACCGGGTTTAAACCCAGCTTTGTTTAAACGGCGCGCCGG0007502550>90PstIGCTGCAGCTGCACTGCAGTGCAAACTGCAGAACCAATGCATTGGAAAACTGCAGCCAATGCATTGGAACTGCAGAACCAATGCATTGGATGCAT010>90>900010>90>900PvuICCGATCGGATCGATCGATTCGCGATCGCGA010002510SacICGAGCTCG1010SacIIGCCGCGGCTCCCCGCGGGGA0500>90SalI

6、GTCGACGTCAAAAGGCCATAGCGGCCGCGCGTCGACGTCTTGGCCATAGCGGCCGCGGACGCGTCGACGTCGGCCATAGCGGCCGCGGAA0101005075ScaIGAGTACTCAAAAGTACTTTT10752575SmaICCCGGGCCCCGGGGCCCCCGGGGGTCCCCCGGGGGA0010>90101050>90SpeIGACTAGTCGGACTAGTCCCGGACTAGTCCGCTAGACTAGTCTAG101000>90>905050SphIGGCATGCCCATGCATGCATGACATGCATGCATGT

7、001002550StuIAAGGCCTTGAAGGCCTTCAAAAGGCCTTTT>90>90>90>90>90>90XbaICTCTAGAGGCTCTAGAGCTGCTCTAGAGCACTAGTCTAGACTAG0>9075750>90>90>90XhoICCTCGAGGCCCTCGAGGGCCGCTCGAGCGG0101002575XmaICCCCGGGGCCCCCGGGGGCCCCCCGGGGGGTCCCCCCGGGGGGA02550>90075>90>90注释：1．如果要加在序列的5‘端，就在酶切位点识