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
切割率% 酶 寡核苷酸序列 2 hr Acc I 20 hr GGTCGACC CGGTCGACCG CCGGTCGACCGG 0 0 0 0 >90 >90 >90 >90 >90 50 >90 >90 10 >90 >90 0 75 25 0 0 50 0 0 25 25 0 0 >90 50 0 0 0 0 >90 >90 >90 >90 >90 >90 >90 >90 25 >90 >90 0 >90 >90 0 0 >90 10 0 50 >90 0 0 >90 50 Afl III CACATGTG CCACATGTGG CCCACATGTGGG Asc I GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA Ava I CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA BamH I CGGATCCG CGGGATCCCG CGCGGATCCGCG Bgl II CAGATCTG GAAGATCTTC GGAAGATCTTCC BssH II GGCGCGCC AGGCGCGCCT TTGGCGCGCCAA BstE II BstX I GGGT(A/T)ACCC AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT Cla I CATCGATG GATCGATC CCATCGATGG CCCATCGATGGG
EcoR I GGAATTCC CGGAATTCCG CCGGAATTCCGG >90 >90 >90 >90 >90 >90 0 0 10 0 >90 >90 0 25 0 50 0 0 0 0 75 75 0 10 10 >90 >90 >90 >90 >90 >90 0 0 75 0 >90 >90 0 50 0 75 0 0 0 0 >90 >90 0 25 50 Hae III GGGGCCCC AGCGGCCGCT TTGCGGCCGCAA Hind III CAAGCTTG CCAAGCTTGG CCCAAGCTTGGG Kpn I GGGTACCC GGGGTACCCC CGGGGTACCCCG Mlu I GACGCGTC CGACGCGTCG Nco I CCCATGGG CATGCCATGGCATG Nde I CCATATGG CCCATATGGG CGCCATATGGCG GGGTTTCATATGAAACCC GGAATTCCATATGGAATTCC GGGAATTCCATATGGAATTCCC Nhe I GGCTAGCC CGGCTAGCCG CTAGCTAGCTAG PCR设计引物时酶切位点的保护碱基表2
切割率% 酶 寡核苷酸序列 2 hr Not I 20 hr TTGCGGCCGCAA ATTTGCGGCCGCTTTA AAATATGCGGCCGCTATAAA ATAAGAATGCGGCCGCTAAACTAT AAGGAAAAAAGCGGCCGCAAAAGGAAAA 0 10 10 25 25 10 >90 0 10 10 90 >90 >90 >90 Nsi I TGCATGCATGCA CCAATGCATTGGTTCTGCAGTT
Pac I TTAATTAA GTTAATTAAC CCTTAATTAAGG 0 0 0 0 0 0 75 0 10 >90 >90 0 0 10 0 10 0 50 0 10 10 0 25 >90 0 25 50 >90 0 10 >90 >90 0 0 25 10 10 0 >90 0 50 75 Pme I GTTTAAAC GGTTTAAACC GGGTTTAAACCC AGCTTTGTTTAAACGGCGCGCCGG Pst I GCTGCAGC TGCACTGCAGTGCA AACTGCAGAACCAATGCATTGG AAAACTGCAGCCAATGCATTGGAA CTGCAGAACCAATGCATTGGATGCAT Pvu I CCGATCGG ATCGATCGAT TCGCGATCGCGA Sac I Sac II CGAGCTCG GCCGCGGC TCCCCGCGGGGA Sal I GTCGACGTCAAAAGGCCATAGCGGCCGC GCGTCGACGTCTTGGCCATAGCGGCCGCGG ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA Sca I GAGTACTC AAAAGTACTTTT 10 75 0 0 10 >90 10 10 0 0 0 0 10 25 75 10 10 50 >90 >90 >90 50 50 0 25 50 Sma I CCCGGG CCCCGGGG CCCCCGGGGG TCCCCCGGGGGA Spe I GACTAGTC GGACTAGTCC CGGACTAGTCCG CTAGACTAGTCTAG Sph I GGCATGCC CATGCATGCATG ACATGCATGCATGT
Stu I AAGGCCTT GAAGGCCTTC AAAAGGCCTTTT >90 >90 >90 0 >90 75 75 0 10 10 0 25 50 >90 >90 >90 >90 0 >90 >90 >90 0 25 75 0 75 >90 >90 Xba I CTCTAGAG GCTCTAGAGC TGCTCTAGAGCA CTAGTCTAGACTAG Xho I CCTCGAGG CCCTCGAGGG CCGCTCGAGCGG Xma I CCCCGGGG CCCCCGGGGG CCCCCCGGGGGG TCCCCCCGGGGGGA
pGEM3-H2a-614-HindIII-R:
CAAGCTT GGCTACCGTGACACAACTCTTT pGEM3-H2a-614-86bp-EcoRI-F:
G GAATTC CAAGTGAGGCCACTGCTCGC pGEM3-H2a-614-142bp-EcoRI-F:
G GAATTC AACATCCAGGCCGTGCTGCT pGEM3-H2a-614-196bp-EcoRI-F:
G GAATTC AGGAGCTCAACAAGCTGCTG pGEM3-H2a-614-86bp-EcoRI-F Mature 108bp Pre 142bp 198bp 252bp Cleavge Vector 34bp 34bp 34bp 22bp 22bp 22bp pGEM3-H2a-614-142bp-EcoRI-F 164bp pGEM3-H2a-614-196bp-EcoRI-F 218bp Pcdna3.0-H2a-614-EcoRI-R:
G GAATTC GGCTACCGTGACACAACTCTTT PCDNA3.0- H2a-614-86bp- HindIII-F:
CAAGCTT CAAGTGAGGCCACTGCTCGC pcna3.0-H2a-614-142bp- HindIII -F:
CAAGCTT AACATCCAGGCCGTGCTGCT pcna3.0-H2a-614-196bp- HindIII -F:
CAAGCTT AGGAGCTCAACAAGCTGCTG