我设计了几对引物,帮我看看!
丁香园论坛
980
tg tctgggtctt gcacaatgac
119521 aatgggtccc tgtttttccc agaggctctt ttgttctgca gggattgaag acactccagt
119581 cccacagtcc ccagctcccc tggggcaggg ttggcagaat ttcgacaaca catttttcca
119641 ccctgactag gatgtgctcc tcatggcagc tgggaaccac tgtccaataa gggcctgggc
119701 ttacacagct gcttctcatt gagttacacc cttaataaaa taatcccatt ttatcctttt
119761 tgtctctctg tcttcctctc tctctgcctt tcctcttctc tctcctcctc tctcatctcc
119821 aggtgcaaat agtctacaaa ccagttgacc tgagcaaggt gacctccaag tgtggctcat
119881 taggcaacat ccatcataaa ccag
目的:克隆这个片断。
结果:
1, 5 TGTCTGGGTCTTGCACAATGACA 3
3 TCCGTTGTAGGTAGTATTTGGTC 5
rating L ength Tm GC% oD Ta opt
sense 88 23 63.5 47.8 32.7 1
Anti-sense 100 23 60.8 43.5 33.6 1
product 82 406 88.2 49.0 55.0
HairpinΔG=0.1
2. 5 TGTCTGGGTCTTGCACAATGAC 3
3 TCCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 22 60.6 50 33.4 1
Anti-sense 100 23 60.8 43.5 33.6 1
product 86 406 88.2 49.0 54.9
Hairpin ΔG=0.3
3. 5 TGTCTGGGTCTTGCACAAT 3
3 CGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 19 53.7 47.4 33.8 1
Anti-sense 100 21 55.5 42.9 33.1 1
product 84 40 6 88.2 49.0 52.8
Hairpin ΔG=0.2
4. 5 TGTCTGGGTCTTGCACAATGACA 3
3 CCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 23 63.5 47.8 32.7 1
Anti-sense 100 22 59.6 45.5 33.4 1
product 82 406 88.2 49.0 54.6
Hairpin ΔG=-0.1
5 5 TGTCTGGGTCTTGCACAATGAC 3
3 CCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 22 60.6 50 33.4 1
Anti-sense 100 22 59.6 45.5 33.4 1
product 84 406 88.2 49.0 54.6
Hairpin ΔG=0.3
后面两对有连续的CC,影响大吗?
ΔG的正负是什么意思?对结果有何影响?
119521 aatgggtccc tgtttttccc agaggctctt ttgttctgca gggattgaag acactccagt
119581 cccacagtcc ccagctcccc tggggcaggg ttggcagaat ttcgacaaca catttttcca
119641 ccctgactag gatgtgctcc tcatggcagc tgggaaccac tgtccaataa gggcctgggc
119701 ttacacagct gcttctcatt gagttacacc cttaataaaa taatcccatt ttatcctttt
119761 tgtctctctg tcttcctctc tctctgcctt tcctcttctc tctcctcctc tctcatctcc
119821 aggtgcaaat agtctacaaa ccagttgacc tgagcaaggt gacctccaag tgtggctcat
119881 taggcaacat ccatcataaa ccag
目的:克隆这个片断。
结果:
1, 5 TGTCTGGGTCTTGCACAATGACA 3
3 TCCGTTGTAGGTAGTATTTGGTC 5
rating L ength Tm GC% oD Ta opt
sense 88 23 63.5 47.8 32.7 1
Anti-sense 100 23 60.8 43.5 33.6 1
product 82 406 88.2 49.0 55.0
HairpinΔG=0.1
2. 5 TGTCTGGGTCTTGCACAATGAC 3
3 TCCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 22 60.6 50 33.4 1
Anti-sense 100 23 60.8 43.5 33.6 1
product 86 406 88.2 49.0 54.9
Hairpin ΔG=0.3
3. 5 TGTCTGGGTCTTGCACAAT 3
3 CGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 19 53.7 47.4 33.8 1
Anti-sense 100 21 55.5 42.9 33.1 1
product 84 40 6 88.2 49.0 52.8
Hairpin ΔG=0.2
4. 5 TGTCTGGGTCTTGCACAATGACA 3
3 CCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 23 63.5 47.8 32.7 1
Anti-sense 100 22 59.6 45.5 33.4 1
product 82 406 88.2 49.0 54.6
Hairpin ΔG=-0.1
5 5 TGTCTGGGTCTTGCACAATGAC 3
3 CCGTTGTAGGTAGTATTTGGTC 5
rating length Tm GC% oD Ta opt
sense 88 22 60.6 50 33.4 1
Anti-sense 100 22 59.6 45.5 33.4 1
product 84 406 88.2 49.0 54.6
Hairpin ΔG=0.3
后面两对有连续的CC,影响大吗?
ΔG的正负是什么意思?对结果有何影响?