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2008116068木蟲(chóng) (著名寫(xiě)手)
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[求助]
PCR擴(kuò)增出現(xiàn)的問(wèn)題
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我從一個(gè)8K質(zhì)粒上擴(kuò)增一段2.8K的目的片段，無(wú)論自己設(shè)計(jì)引物（16到22個(gè)堿基）還是按文獻(xiàn)提供的引物（35和38個(gè)堿基），均擴(kuò)增不出來(lái)。已經(jīng)花了一個(gè)月的時(shí)間了，如果再擴(kuò)增不出來(lái)，下個(gè)月就要換課題了。 目的片度的特點(diǎn)是GC含量約30%。 各位蟲(chóng)友，有什么好的建議嗎？對(duì)我來(lái)說(shuō)很重要，就麻煩大家了。 謝謝  |
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天使托
專家顧問(wèn) (職業(yè)作家)
T.Shen
-
專家經(jīng)驗(yàn): +57 - MicEPI: 13
- 應(yīng)助: 95 (初中生)
- 貴賓: 0.263
- 金幣: 16332.8
- 散金: 593
- 紅花: 74
- 沙發(fā): 9
- 帖子: 3652
- 在線: 289小時(shí)
- 蟲(chóng)號(hào): 441757
- 注冊(cè): 2007-10-27
- 專業(yè): 微生物生理與生物化學(xué)
- 管轄: 微生物
2樓2011-06-14 17:48:11
3樓2011-06-14 18:41:43
2008116068
木蟲(chóng) (著名寫(xiě)手)
- 應(yīng)助: 3 (幼兒園)
- 金幣: 4612.6
- 散金: 294
- 紅花: 3
- 帖子: 1115
- 在線: 364.5小時(shí)
- 蟲(chóng)號(hào): 701164
- 注冊(cè): 2009-02-14
- 性別: GG
- 專業(yè): 微生物生理與生物化學(xué)
|
目標(biāo)序列為 CTAACTTATAGGGGTAACACTTAAAAAAGAATCAATAACGATAGAAACCGCTCCTAAAGCAGGTGCATTTTTTCCTAACGAAGAAGGCAATAGTTCACATTTATTGTCTAAATGAGAATGGACTCTAGAAGAAACTTCGTTTTTAATCGTATTTAAAACAATGGGATGAGATTCAATTATATGATTTCTCAAGATAACAGCTTCTATATCAAATGTATTAAGGATATTGGTTAATCCAATTCCGATATAAAAGCCAAAGTTTTGAAGTGCATTTAACATTTCTACATCATTTTTATTTGCGCGTTCCACAATCTCTTTTCGAGAAATATTCTTTTCTTCTTTAGAGAGCGAAGCCAGTAACGCTTTTTCAGAAGCATATAATTCCCAACAGCCTCGATTTCCACAGCTGCATTTGGGTCCATTAAAATCTATCGTCATATGACCCATTTCCCCAGAAAAACCCTGAACACCTTTATACAATTCGTTGTTAATAACAAGTCCAGTTCCAATTCCGATATTAATACTGATGTAAACGATGTTTTCATAGTTTTTTGTCATACCAAATACTTTTTCACCGTATGCTCCTGCATTAGCTTCATTTTCAACAAAAACCGGAACATTAAACTCACTCTCAATTAAAAACTGCAAATCTTTGATATTCCAATTTAAGTTAGGCATGAAAATAATTTGCTGATGACGATCTACAAGGCCTGGAACACAAATTCCTATTCCGACTAGACCATAAGGGGACTCAGGCATATGGGTTACAAAACCATGAATAAGTGCAAATAAAATCTCTTTTACTTCACTAGCGGAAGAACTAGACAAGTCAGAAGTCTTCTCGAGAATAATATTTCCTTCTAAGTCGGTTAGAATTCCGTTAAGATAGTCGACTCCTATATCAATACCAATCGAGTAGCCTGCATTCTTATTAAAAACAAGCATTACAGGTCTTCTGCCGCCTCTAGATTGCCCTGCCCCAATTTCAAAAATAAAATCTTTTTCAAGCAGTGTATTTACTTGAGAGGAGACAGTAGACTTGTTTAATCCTGTAATCTCAGAGAGAGTTGCCCTGGAGACAGGGGAGTTCTTCAAAATTTCATCTAATATTAATTTTTGATTCATTTTTTTTACTAAAGCTTGATCTGCAATTTGAATAATAACCACTCCTTTGTTTATCCACCGAACTAAGTTGGTGTTTTTTGAAGCTTGAATTAGATATTTAAAAGTATCATATCTAATATTATAACTAAATTTTCTAAAAAAAACATTGAAATAAACATTTATTTTGTATATGATGAGATAAAGTTAGTTTATTGGATAAACAAACTAACTCAATTAAGATAGTTGATGGATAAACTTGTTCACTTAAATCAAAGGGGGAAATGACAAATGGTCCAAACTAGTGATATCTAAAAATCAAAGGGGGAAATGAGATCTATGGTAAGCCGATACGTACCCGATATGGGCGATCTGATTTGGGTTGATTTTGACCCGACAAAAGGTAGCGAGCAAGCTGGACATCGTCCAGCTGTTGTCCTGAGTCCTTTCATGTACAACAACAAAACAGGTATGTGTCTGTGTGTTCCTTGTACAACGCAATCAAAAGGATATCCGTTCGAAGTTGTTTTATCCGGTCAGGAACGTGATGGCGTAGCGTTAGCTGATCAGGTAAAAAGTATCGCCTGGCGGGCAAGAGGAGCAACGAAGAAAGGAACAGTTGCCCCAGAGGAATTACAACTCATTAAAGCCAAAATTAACGTACTGATTGGGTAGGCATGCTAATGCGGCCGCGAATTCCTGCAGCCCTGGCGAATGGCGATTTTCGTTCGTGAATACATGTTATAATAACTATAACTAATAACGTAACGTGACTGGCAAGAGATATTTTTAAAACAATGAATAGGTTTACACTTACTTTAGTTTTATGGAAATGAAAGATCATATCATATATAATCTAGAATAAAATTAACTAAAATAATTATTATCTAGATAAAAAATTTAGAAGCCAATGAAATCTATAAATAAACTAAATTAAGTTTATTTAATTAACAACTATGGATATAAAATAGGTACTAATCAAAATAGTGAGGAGGATATATTTGAATACATACGAACAAATTAATAAAGTGAAAAAAATACTTCGGAAACATTTAAAAAATAACCTTATTGGTACTTACATGTTTGGATCAGGAGTTGAGAGTGGACTAAAACCAAATAGTGATCTTGACTTTTTAGTCGTCGTATCTGAACCATTGACAGATCAAAGTAAAGAAATACTTATACAAAAAATTAGACCTATTTCAAAAAAAATAGGAGATAAAAGCAACTTACGATATATTGAATTAACAATTATTATTCAGCAAGAAATGGTACCGTGGAATCATCCTCCCAAACAAGAATTTATTTATGGAGAATGGTTACAAGAGCTTTATGAACAAGGATACATTCCTCAGAAGGAATTAAATTCAGATTTAACCATAATGCTTTACCAAGCAAAACGAAAAAATAAAAGAATATACGGAAATTATGACTTAGAGGAATTACTACCTGATATTCCATTTTCTGATGTGAGAAGAGCCATTATGGATTCGTCAGAGGAATTAATAGATAATTATCAGGATGATGAAACCAACTCTATATTAACTTTATGCCGTATGATTTTAACTATGGACACGGGTAAAATCATACCAAAAGATATTGCGGGAAATGCAGTGGCTGAATCTTCTCCATTAGAACATAGGGAGAGAATTTTGTTAGCAGTTCGTAGTTATCTTGGAGAGAATATTGAATGGACTAATGAAAATGTAAATTTAACTATAAACTATTTAAATAACAGATTAAAAAAATTATAAAAAAATTGAAAAAATGGTGGAAACACTTTTTTCAATTTTTTTGTTTTATTATTTAATATTTGGGAAATATTCATTCTAATTGGTAATCAGATTTTAGAAAACAATAAACCCTTGCATAGGGGGATCTCG |
4樓2011-06-14 22:44:11
2008116068
木蟲(chóng) (著名寫(xiě)手)
- 應(yīng)助: 3 (幼兒園)
- 金幣: 4612.6
- 散金: 294
- 紅花: 3
- 帖子: 1115
- 在線: 364.5小時(shí)
- 蟲(chóng)號(hào): 701164
- 注冊(cè): 2009-02-14
- 性別: GG
- 專業(yè): 微生物生理與生物化學(xué)
5樓2011-06-14 22:48:00
lovelyxixi81
金蟲(chóng) (小有名氣)
- 應(yīng)助: 0 (幼兒園)
- 金幣: 3095.1
- 帖子: 289
- 在線: 138.3小時(shí)
- 蟲(chóng)號(hào): 866243
- 注冊(cè): 2009-10-09
- 專業(yè): 微生物生理與生物化學(xué)
6樓2011-06-14 23:29:19
2008116068
木蟲(chóng) (著名寫(xiě)手)
- 應(yīng)助: 3 (幼兒園)
- 金幣: 4612.6
- 散金: 294
- 紅花: 3
- 帖子: 1115
- 在線: 364.5小時(shí)
- 蟲(chóng)號(hào): 701164
- 注冊(cè): 2009-02-14
- 性別: GG
- 專業(yè): 微生物生理與生物化學(xué)
|
載體全序列 1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga 61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt 121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat 181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt 241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg 301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg 361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg 421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa 481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat 541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca 601 catggaaaca cacaaattaa aaactggtct gatcggatcc ctaacttata ggggtaacac 661 ttaaaaaaga atcaataacg atagaaaccg ctcctaaagc aggtgcattt tttcctaacg 721 aagaaggcaa tagttcacat ttattgtcta aatgagaatg gactctagaa gaaacttcgt 781 ttttaatcgt atttaaaaca atgggatgag attcaattat atgatttctc aagataacag 841 cttctatatc aaatgtatta aggatattgg ttaatccaat tccgatataa aagccaaagt 901 tttgaagtgc atttaacatt tctacatcat ttttatttgc gcgttccaca atctcttttc 961 gagaaatatt cttttcttct ttagagagcg aagccagtaa cgctttttca gaagcatata 1021 attcccaaca gcctcgattt ccacagctgc atttgggtcc attaaaatct atcgtcatat 1081 gacccatttc cccagaaaaa ccctgaacac ctttatacaa ttcgttgtta ataacaagtc 1141 cagttccaat tccgatatta atactgatgt aaacgatgtt ttcatagttt tttgtcatac 1201 caaatacttt ttcaccgtat gctcctgcat tagcttcatt ttcaacaaaa accggaacat 1261 taaactcact ctcaattaaa aactgcaaat ctttgatatt ccaatttaag ttaggcatga 1321 aaataatttg ctgatgacga tctacaaggc ctggaacaca aattcctatt ccgactagac 1381 cataagggga ctcaggcata tgggttacaa aaccatgaat aagtgcaaat aaaatctctt 1441 ttacttcact agcggaagaa ctagacaagt cagaagtctt ctcgagaata atatttcctt 1501 ctaagtcggt tagaattccg ttaagatagt cgactcctat atcaatacca atcgagtagc 1561 ctgcattctt attaaaaaca agcattacag gtcttctgcc gcctctagat tgccctgccc 1621 caatttcaaa aataaaatct ttttcaagca gtgtatttac ttgagaggag acagtagact 1681 tgtttaatcc tgtaatctca gagagagttg ccctggagac aggggagttc ttcaaaattt 1741 catctaatat taatttttga ttcatttttt ttactaaagc ttgatctgca atttgaataa 1801 taaccactcc tttgtttatc caccgaacta agttggtgtt ttttgaagct tgaattagat 1861 atttaaaagt atcatatcta atattataac taaattttct aaaaaaaaca ttgaaataaa 1921 catttatttt gtatatgatg agataaagtt agtttattgg ataaacaaac taactcaatt 1981 aagatagttg atggataaac ttgttcactt aaatcaaagg gggaaatgac aaatggtcca 2041 aactagtgat atctaaaaat caaaggggga aatgagatct atggtaagcc gatacgtacc 2101 cgatatgggc gatctgattt gggttgattt tgacccgaca aaaggtagcg agcaagctgg 2161 acatcgtcca gctgttgtcc tgagtccttt catgtacaac aacaaaacag gtatgtgtct 2221 gtgtgttcct tgtacaacgc aatcaaaagg atatccgttc gaagttgttt tatccggtca 2281 ggaacgtgat ggcgtagcgt tagctgatca ggtaaaaagt atcgcctggc gggcaagagg 2341 agcaacgaag aaaggaacag ttgccccaga ggaattacaa ctcattaaag ccaaaattaa 2401 cgtactgatt gggtaggcat gctaatgcgg ccgcgaattc ctgcagccct ggcgaatggc 2461 gattttcgtt cgtgaataca tgttataata actataacta ataacgtaac gtgactggca 2521 agagatattt ttaaaacaat gaataggttt acacttactt tagttttatg gaaatgaaag 2581 atcatatcat atataatcta gaataaaatt aactaaaata attattatct agataaaaaa 2641 tttagaagcc aatgaaatct ataaataaac taaattaagt ttatttaatt aacaactatg 2701 gatataaaat aggtactaat caaaatagtg aggaggatat atttgaatac atacgaacaa 2761 attaataaag tgaaaaaaat acttcggaaa catttaaaaa ataaccttat tggtacttac 2821 atgtttggat caggagttga gagtggacta aaaccaaata gtgatcttga ctttttagtc 2881 gtcgtatctg aaccattgac agatcaaagt aaagaaatac ttatacaaaa aattagacct 2941 atttcaaaaa aaataggaga taaaagcaac ttacgatata ttgaattaac aattattatt 3001 cagcaagaaa tggtaccgtg gaatcatcct cccaaacaag aatttattta tggagaatgg 3061 ttacaagagc tttatgaaca aggatacatt cctcagaagg aattaaattc agatttaacc 3121 ataatgcttt accaagcaaa acgaaaaaat aaaagaatat acggaaatta tgacttagag 3181 gaattactac ctgatattcc attttctgat gtgagaagag ccattatgga ttcgtcagag 3241 gaattaatag ataattatca ggatgatgaa accaactcta tattaacttt atgccgtatg 3301 attttaacta tggacacggg taaaatcata ccaaaagata ttgcgggaaa tgcagtggct 3361 gaatcttctc cattagaaca tagggagaga attttgttag cagttcgtag ttatcttgga 3421 gagaatattg aatggactaa tgaaaatgta aatttaacta taaactattt aaataacaga 3481 ttaaaaaaat tataaaaaaa ttgaaaaaat ggtggaaaca cttttttcaa tttttttgtt 3541 ttattattta atatttggga aatattcatt ctaattggta atcagatttt agaaaacaat 3601 aaacccttgc atagggggat ctcgacatgg atgagcgatg atgatatccg tttaggctgg 3661 gcggtgatag cttctcgttc aggcagtacg cctcttttct tttccagacc tgagggaggc 3721 ggaaatggtg tgaggttccc ggggaaaagc caaataggcg atcgcgggag tgctttattt 3781 gaagatcagg ctatcactgc ggtcaataga tttcacaatg tgatggctgg acagcctgag 3841 gaactctcga acccgaatgg aaacaaccag atatttatga atcagcgcgg ctcacatggc 3901 gttgtgctgg caaatgcagg ttcatcctct gtctctatca atacggcaac aaaattgcct 3961 gatggcaggt atgacaataa agctggagcg ggttcatttc aagtgaacga tggtaaactg 4021 acaggcacga tcaatgccag gtctgtagct gtgctttatc ctgatgatat tgcaaaagcg 4081 cctcatgttt tccttgagaa ttacaaaaca ggtgtaacac attctttcaa tgatcaactg 4141 acgattacct tgcgtgcaga tgcgaataca acaaaagccg tttatcaaat caataatgga 4201 ccagacgaca ggcgtttaag gatggagatc aattcacaat cggaaaagga gatccaattt 4261 ggcaaaacat acaccatcat gttaaaagga acgaacagtg atggtgtaac gaggaccgag 4321 aaatacagtt ttgttaaaag agatccagcg tcggccaaaa ccatcggcta tcaaaatccg 4381 aatcattgga gccaggtaaa tgcttatatc tataaacatg atgggagccg agtaattgaa 4441 ttgaccggat cttggcctgg aaaaccaatg actaaaaatg cagacggaat ttacacgctg 4501 acgctgcctg cggacacgga tacaaccaac gcaaaagtga tttttaataa tggcagcgcc 4561 caagtgcccg gtcagaatca gcctggcttt gattacgtgc taaatggttt atataatgac 4621 tcgggcttaa gcggttctct tccccattga gggcaaggct agacgggact taccgaaaga 4681 aaccatcaat gatggtttct tttttgttca taaatcagac aaaacttttc tcttgcaaaa 4741 gtttgtgaag tgttgcacaa tataaatgtg aaatacttca caaacaaaaa gacatcaaag 4801 agaaacatac cctgcaagga tgctgatatt gtctgcattt gcgccggagc aaaccaaaaa 4861 cctggtgaga cacgccttga attagtagaa aagaacttga agattttcaa aggcatcgtt 4921 agtgaagtca tggcgagcgg atttgacggc attttcttag tcgcgacgcg aggctggatg 4981 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc 5041 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct 5101 cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta tgccgcctcg 5161 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc 5221 cccgcgttgc gtcgcggtgc atggagccgg gccacctact gaagtggatt tctttaagag 5281 ctccgataac cagaagcaat aaaaaatcaa atcggatttc actatataat ctcactttat 5341 ctaagatgaa tccgatggaa gcatcctgtt ttctctcaat ttttttatct aaaacccagc 5401 gttcgatgct tctttgagcg aacgatcaaa aataagtgcc ttcccatcaa aaaaatattc 5461 tcaacataaa aaactttgtg taatacttgt aacgctacat ggagattaac tcaatctaga 5521 gggtattaat aatgatccac agtagcgtaa agcgttgggg aaattcaccg gcggtgcgga 5581 tcccggctac gttaatgcag gcgctcaatc tgaatattga tgatgaagtg aagattgacc 5641 tggtggatgg caaattaatt attgagccag tgcgtaaaga gcccgtattt acgcttgctg 5701 aactggtcaa cgacatcacg ccggaaaacc tccacgagaa tatcgactgg ggagagccga 5761 aagataagga agtctggtaa atcgattatg tcttttgcgc agtcggctta aaccagtttt 5821 cgctggtgcg aaaaaagagt gtcttgtgac acctaaattc aaaatctatc ggtcagattt 5881 ataccgattt gattttatat attcttgaat aacatacgcc gagttatcac ataaaagcgg 5941 gaaccaatca tcaaatttaa acttcattgc ataatccatt aaactcttaa attctacgat 6001 tccttgttca tcaataaact caatcatttc tttaattaat ttatatctat ctgttgttgt 6061 tttctttaat aattcatcaa catctacacc gccataaact atcatatctt ctttttgata 6121 tttaaattta ttaggatctt aaggcctagg tctagagtct ttgttttgac gccattagcg 6181 tacgtaacaa tcctcgttaa aggacaagga cctgagcgga agtgtatcgt acagtagacg 6241 gagtatacta gtatagtcta tagtccgtgg aattattata tttatctccg acgatattct 6301 catcagtgaa atccagctgg agttctttag caaatttttt tattagctga acttagtatt 6361 agtggccata ctcctccaat ccaaagctat ttagaaagat tactatatcc tcaaacaggc 6421 ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag catcgccggc atgtccccct 6481 ggcggacggg aagtatccag ctcgaggtcg ggccgcgttg ctggcgtttt tccataggct 6541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 6601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 6661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 6721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 6781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 6841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 6901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 6961 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 7021 agttgatagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 7081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 7141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 7201 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 7261 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 7321 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 7381 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 7441 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 7501 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 7561 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc 7621 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 7681 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 7741 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 7801 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 7861 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc 7921 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 7981 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 8041 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 8101 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 8161 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 8221 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 8281 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc 8341 ctttcgtctt caagaatt |
7樓2011-06-15 09:30:02
2008116068
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8樓2011-06-15 15:13:36
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好吧，這么說(shuō)吧： 1。確保你的模板沒(méi)有任何問(wèn)題，質(zhì)粒用試劑盒重新抽提。 2.你要擴(kuò)增的片段是從640-3490，AT含量較高，引物不好設(shè)計(jì)，你應(yīng)該考試試專門(mén)針對(duì)AT-rich的擴(kuò)增方法。 3.如果還是不行的話，建議你不要再糾結(jié)于引物擴(kuò)增，而改用酶切回收，看了一下酶切位點(diǎn)還是有很多可以選擇的，提供兩個(gè)和你兩端比較接近的 5‘端： BamHI 2 G/GATCC 635 5578 3'端：SwaI 2 ATTT/AAAT 3470 6123 直接回收635-3470片段大小的， 后面看你想干嘛了，用Taq酶補(bǔ)平加A尾進(jìn)行TA克��；或者再用你的引物PCR試試，應(yīng)該沒(méi)有什么太麻煩的二級(jí)結(jié)構(gòu)了 都試試吧 |
9樓2011-06-18 16:08:37
10樓2011-06-18 16:10:43
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