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_1714116324592_0引物时酶切位点的保护酶寡核苷酸序列切割率%2hr20hrAccIGGTCGACC00CGGTCGACCG00CCGGTCGACCGG00AflIIICACATGTG00CCACATGTGG>90>90CCCACATGTGGG>90>90AscIGGCGCGCC>90>90AGGCGCGCCT>90>90TTGGCGCGCCAA>90>90AvaICCCCGGGG50>90CCCCCGGGGG>90>90TCCCCCGGGGGA>90>90BamHICGGATCCG1025CGGGATCCCG>90>90CGCGGATCCGCG>90>90BglIICAGATCTG00GAAGATCTTC75>90GGAAGATCTTCC25>90BssHIIGGCGCGCC00AGGCGCGCCT00TTGGCGCGCCAA50>90BstEIIGGGT(A/T)ACCC010BstXIAACTGCAGAACCAATGCATTGG00AAAACTGCAGCCAATGCATTGGAA2550CTGCAGAACCAATGCATTGGATGCAT25>90ClaICATCGATG00GATCGATC00CCATCGATGG>90>90CCCATCGATGGG5050EcoRIGGAATTCC>90>90CGGAATTCCG>90>90CCGGAATTCCGG>90>90HaeIIIHindIIIKpnIMluINcoINdeINheINotINsiIPacIPmeIGGGGCCCC>90>90AGCGGCCGCT>90>90TTGCGGCCGCAA>90>90CAAGCTTG00CCAAGCTTGG00CCCAAGCTTGGG1075GGGTACCC00GGGGTACCCC>90>90CGGGGTACCCCG>90>90GACGCGTC00CGACGCGTCG2550CCCATGGG00CATGCCATGGCATG5075CCATATGG00CCCATATGGG00CGCCATATGGCG00GGGTTTCATATGAAACCC00GGAATTCCATATGGAATTCC75>90GGGAATTCCATATGGAATTCCC75>90GGCTAGCC00CGGCTAGCCG1025CTAGCTAGCTAG1050TTGCGGCCGCAA00ATTTGCGGCCGCTTTA1010AAATATGCGGCCGCTATAAA1010ATAAGAATGCGGCCGCTAAACTAT2590AAGGAAAAAAGCGGCCGCAAAAGGAAAA25>90TGCATGCATGCA10>90CCAATGCATTGGTTCTGCAGTT>90>90TTAATTAA00GTTAATTAAC025CCTTAATTAAGG0>90GTTTAAAC00GGTTTAAACC025GGGTTTAAACCC050AGCTTTGTTTAAACGGCGCGCCGG75>90PstIPvuISacISacIISalIScaISmaISpeISphIStuIXbaIXhoIGCTGCAGC00TGCACTGCAGTGCA1010AACTGCAGAACCAATGCATTGG>90>90AAAACTGCAGCCAATGCATTGGAA>90>90CTGCAGAACCAATGCATTGGATGCAT00CCGATCGG00ATCGATCGAT1025TCGCGATCGCGA010CGAGCTCG1010GCCGCGGC00TCCCCGCGGGGA50>90GTCGACGTCAAAAGGCCATAGCGGCCGC00GCGTCGACGTCTTGGCCATAGCGGCCGCGG1050ACGCGTCGACGTCGGCCATAGCGGCCGCG1075GAAGAGTACTC1025AAAAGTACTTTT7575CCCGGG010CCCCGGGG010CCCCCGGGGG1050TCCCCCGGGGGA>90>90GACTAGTC10>90GGACTAGTCC10>90CGGACTAGTCCG050CTAGACTAGTCTAG050GGCATGCC00CATGCATGCATG025ACATGCATGCATGT1050AAGGCCTT>90>90GAAGGCCTTC>90>90AAAAGGCCTTTT>90>90CTCTAGAG00GCTCTAGAGC>90>90TGCTCTAGAGCA75>90CTAGTCTAGACTAG75>90CCTCGAGG00CCCTCGAGGG1025CCGCTCGAGCGG1075XmaI说明：CCCCGGGG00CCCCCGGGGG2575CCCCCCGGGGGG50>90TCCCCCCGGGGGGA>90>901．如果要加在序列的5‘端，就在酶切位点辨别碱基序列（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，就在酶切位点辨别碱基序列（红色）的3’端加上相应的碱基（黑色）。2．切割率：正确辨别并酶切的效率3。加保护碱基时最好采用切割率高时加的相应碱基。
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