请大家分析一下我这引物是否可行----急!
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我要对大鼠的IL-18RβmRNA进行RT-PCR现设计了如下两条引物:
引物对1:扩增区域 1~522 退火温度59℃。
F1 ATGCTTAAATCCAGATGTGGCG
R1 CCCATAAAACAGGGCTACCTCC
引物对2:扩增区域600~1113 退火温度58℃
F2 AGGCTTGTACGTGTGCGATTAC
R2 TTCAATCCAGTGCCAGTAGAGC
请各位高手帮我分析分析这两对引物怎样,替我指点指点!谢谢!
以下是全系列:
1 atgcttaaat ccagatgtgg cgatgatgat ggtgaacagc catttcaagc ccatgaagga
61 gaacagactg aaaatataaa cccaggaagt cccttcagaa cagcccggga aaagctcttt
121 gttgggccac aggaactgtg ccagacactt ggaggtgggg aaattcctgc ttctacctgt
181 gagaaggcca gtggaaggga aatccctcaa gtagcagatg agactcttag tcagaaaaag
241 gtttccgagg gaggtatgaa tacagcagga ggaagtggga atattgggct ctgtgggaga
301 gctaagagag gaggaagact gaaggtcagt catggcagct gccctgatgt gtccttaagg
361 aagccgttta cacagatcaa acacattcag tccaccttgg ccttctttcc tgtacctgag
421 ctttactctg catcgtccta cagtgatgtt gattttttaa ttggttcccc tgtgttcttc
481 gcaactctcc tcagactcag ggaggtagcc ctgttttatg ggagccgaga ggatggaaga
541 ctgctgcccg agtataagaa aaatccaatt gagatggagg atatttatga ttttcaccaa
601 ggcttgtacg tgtgcgatta cactcagtca gataatgtga gttcctggac cgtccgagct
661 gtggttcaag tgagaaccat cgatgaggac accaatgtga agccggacat tctggatccc
721 gttacagaca ctctggacgt ggagcttgga aaggctttta ctctaccctg cagagtacag
781 tttggcttcc aaagatattc aaagcctgta ataaagtggt atgtcaaaga atctacacag
841 gagtgggaaa tttcagaatt tgtggagaaa aggattcaat ccactttgaa gaacgaagtc
901 attgaacata ctgtcttctt gagagaagtc acccaaagag atctcagcag aaagtttgtt
961 tgctttgctc agaactccat tgggaacaca acacggacca tccgtctgag gaagaaggaa
1021 ggagtggtgt ttctatacat cctcctcggc atgactgtga tgctggtggg tgtactggtg
1081 gcagctgctt tgctctactg gcactggatt gaagttgtcc tgctgtgccg aaccttcaag
1141 aacaaagatg agactctgag ggataagaag gaatttgatg catttgtatc ctatgcaaat
1201 tggagctctc ctgagactga agccacagga tctctgagtg aggaacactt ggctctaaat
1261 cttttcccgg aagtgctaga gaacacatat ggatacagct tgtgtttgtt tgaaagagac 1321 gtgaccccag ggggagtata tactgacgac attgtgagca tcattaagag aagccgaaga
1381 ggaatattta tcctgagtcc cagatacatc aatggacccc gtgtctttga actacaagca 1441 gcagtgaacc ttgccttggt tgatcagaca ctgaagttga ttttaattaa gttctgttcc
1501 ttccaagagc cagaatctct tccttacctt gtcaaaaagg ccctgcgggt tctccccaca
1561 gtcacgtgga aaggcttgaa gtcggttcac gccacttcta ggttctggac ccaaatacgt
1621 taccacatgc ccgtgaagaa ctccaacaga tttatgttca actggctcag acttatcccg
1681 aagggctttt cccctgagaa ggacctagca acgcagaaac ccttgggagg aatgcccaac
1741 tcgggaaaca gccatggagc tcagaacaga acgcccgatg cagggaacag acacggagct
1801 cagaacctcc ttctctccgg ccaccagaag agaagccgaa gaggaatatt tatcctgagt
1861 cccagataca tcaatggacc ccgtgtcttt gaactacaag cagcagtgaa ccttgccttg
1921 gttgatcaga cactgaagtt gattttaatt aagttctgtt ccttccaaga gccagaatct
1981 cttccttacc ttgtcaaaaa ggccctgcgg gttctcccca cagtcacgtg gaaaggcttg
2041 aagtcggttc acgccacttc taggttctgg acccaaatac gttaccacat gcccgtgaag
2101 aactccaaca gatttatgtt caactggctc agacttatcc cgaagggctt ttcccctgag 2161 aaggacctag caacgcagaa acccttggga ggaatgccca actcgggaaa cagccatgga
2221 gctcagaaca gaacgcccga tgcagggaac agacacggag ctcagaacct ccttctctcc
2281 ggccaccaga agaggtgctg a
引物对1:扩增区域 1~522 退火温度59℃。
F1 ATGCTTAAATCCAGATGTGGCG
R1 CCCATAAAACAGGGCTACCTCC
引物对2:扩增区域600~1113 退火温度58℃
F2 AGGCTTGTACGTGTGCGATTAC
R2 TTCAATCCAGTGCCAGTAGAGC
请各位高手帮我分析分析这两对引物怎样,替我指点指点!谢谢!
以下是全系列:
1 atgcttaaat ccagatgtgg cgatgatgat ggtgaacagc catttcaagc ccatgaagga
61 gaacagactg aaaatataaa cccaggaagt cccttcagaa cagcccggga aaagctcttt
121 gttgggccac aggaactgtg ccagacactt ggaggtgggg aaattcctgc ttctacctgt
181 gagaaggcca gtggaaggga aatccctcaa gtagcagatg agactcttag tcagaaaaag
241 gtttccgagg gaggtatgaa tacagcagga ggaagtggga atattgggct ctgtgggaga
301 gctaagagag gaggaagact gaaggtcagt catggcagct gccctgatgt gtccttaagg
361 aagccgttta cacagatcaa acacattcag tccaccttgg ccttctttcc tgtacctgag
421 ctttactctg catcgtccta cagtgatgtt gattttttaa ttggttcccc tgtgttcttc
481 gcaactctcc tcagactcag ggaggtagcc ctgttttatg ggagccgaga ggatggaaga
541 ctgctgcccg agtataagaa aaatccaatt gagatggagg atatttatga ttttcaccaa
601 ggcttgtacg tgtgcgatta cactcagtca gataatgtga gttcctggac cgtccgagct
661 gtggttcaag tgagaaccat cgatgaggac accaatgtga agccggacat tctggatccc
721 gttacagaca ctctggacgt ggagcttgga aaggctttta ctctaccctg cagagtacag
781 tttggcttcc aaagatattc aaagcctgta ataaagtggt atgtcaaaga atctacacag
841 gagtgggaaa tttcagaatt tgtggagaaa aggattcaat ccactttgaa gaacgaagtc
901 attgaacata ctgtcttctt gagagaagtc acccaaagag atctcagcag aaagtttgtt
961 tgctttgctc agaactccat tgggaacaca acacggacca tccgtctgag gaagaaggaa
1021 ggagtggtgt ttctatacat cctcctcggc atgactgtga tgctggtggg tgtactggtg
1081 gcagctgctt tgctctactg gcactggatt gaagttgtcc tgctgtgccg aaccttcaag
1141 aacaaagatg agactctgag ggataagaag gaatttgatg catttgtatc ctatgcaaat
1201 tggagctctc ctgagactga agccacagga tctctgagtg aggaacactt ggctctaaat
1261 cttttcccgg aagtgctaga gaacacatat ggatacagct tgtgtttgtt tgaaagagac 1321 gtgaccccag ggggagtata tactgacgac attgtgagca tcattaagag aagccgaaga
1381 ggaatattta tcctgagtcc cagatacatc aatggacccc gtgtctttga actacaagca 1441 gcagtgaacc ttgccttggt tgatcagaca ctgaagttga ttttaattaa gttctgttcc
1501 ttccaagagc cagaatctct tccttacctt gtcaaaaagg ccctgcgggt tctccccaca
1561 gtcacgtgga aaggcttgaa gtcggttcac gccacttcta ggttctggac ccaaatacgt
1621 taccacatgc ccgtgaagaa ctccaacaga tttatgttca actggctcag acttatcccg
1681 aagggctttt cccctgagaa ggacctagca acgcagaaac ccttgggagg aatgcccaac
1741 tcgggaaaca gccatggagc tcagaacaga acgcccgatg cagggaacag acacggagct
1801 cagaacctcc ttctctccgg ccaccagaag agaagccgaa gaggaatatt tatcctgagt
1861 cccagataca tcaatggacc ccgtgtcttt gaactacaag cagcagtgaa ccttgccttg
1921 gttgatcaga cactgaagtt gattttaatt aagttctgtt ccttccaaga gccagaatct
1981 cttccttacc ttgtcaaaaa ggccctgcgg gttctcccca cagtcacgtg gaaaggcttg
2041 aagtcggttc acgccacttc taggttctgg acccaaatac gttaccacat gcccgtgaag
2101 aactccaaca gatttatgtt caactggctc agacttatcc cgaagggctt ttcccctgag 2161 aaggacctag caacgcagaa acccttggga ggaatgccca actcgggaaa cagccatgga
2221 gctcagaaca gaacgcccga tgcagggaac agacacggag ctcagaacct ccttctctcc
2281 ggccaccaga agaggtgctg a