紧急求教各位高手:引物设计
丁香园论坛
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我要作半定量RT-PCR,在文献上查引物序列用Primer分析,才50-60分,不知是直接引用文献的好,还是自己设计.下面是我从文献上查的几对引物,请帮忙看一下,可否直接引用,还是要重新设计。本人就要开始作RT-PCR了,急需您的帮助。
Smad3 (308 bp, GenBank Accession No. NM_005902)
5’-CAGAACGTCAACACCAAGT (nt 238–256)
and 5’-ATGGAATGGCTGTAGTCGT (nt 545–527);
Smad3
5’-TCCCCAGCACATAATAACTT-3’
and 3’-TAAAAACAGGTCAGAGGGT-5’
Smad3
上游 TAAGTGAGCAGAACAGGTAGTATT
下游 CTCCCTCCCTCCCCACTCCCAAGTC
Smad2 AF027964
Sense aag aag tca gct ggt ggg t nt 161–179
Antisense gcc tgt tgt atc cca ctg a nt 406–388
Smad2
上游 TAGGTGGGGAAGTTTTTGCTGATG
下游 CGTCTGCCTTCGGTATTCTG
Smad7 AF015261
Sense gcc ctc tct gga tat ctt ct nt 843–862
Antisense gct gca taa act cgt ggt ca nt 1162–1143
Smad7
上游 ACCGCAGCAGTTACCCCATCTT
下游 GGCTACCGGCTGTTGAA
Human connective tissue growth factor (hCTGF), GenBank Accession No. X92511;
sense 5-CTCGCGGCTTACCGACTG-3 (positions663–681);
anti-sense 5-GCACTTGAACTCCACCGC-3 (positions1032–1050);
CTGF forward primer: 5-GAGGAAAACATTAAGAAGGGCAAA-3,
reverse primer: 5-CGGCACAGGTCTTGATGA-3,
Smad3 (308 bp, GenBank Accession No. NM_005902)
5’-CAGAACGTCAACACCAAGT (nt 238–256)
and 5’-ATGGAATGGCTGTAGTCGT (nt 545–527);
Smad3
5’-TCCCCAGCACATAATAACTT-3’
and 3’-TAAAAACAGGTCAGAGGGT-5’
Smad3
上游 TAAGTGAGCAGAACAGGTAGTATT
下游 CTCCCTCCCTCCCCACTCCCAAGTC
Smad2 AF027964
Sense aag aag tca gct ggt ggg t nt 161–179
Antisense gcc tgt tgt atc cca ctg a nt 406–388
Smad2
上游 TAGGTGGGGAAGTTTTTGCTGATG
下游 CGTCTGCCTTCGGTATTCTG
Smad7 AF015261
Sense gcc ctc tct gga tat ctt ct nt 843–862
Antisense gct gca taa act cgt ggt ca nt 1162–1143
Smad7
上游 ACCGCAGCAGTTACCCCATCTT
下游 GGCTACCGGCTGTTGAA
Human connective tissue growth factor (hCTGF), GenBank Accession No. X92511;
sense 5-CTCGCGGCTTACCGACTG-3 (positions663–681);
anti-sense 5-GCACTTGAACTCCACCGC-3 (positions1032–1050);
CTGF forward primer: 5-GAGGAAAACATTAAGAAGGGCAAA-3,
reverse primer: 5-CGGCACAGGTCTTGATGA-3,