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| 5 | 1/1 | 返回列表 |
| 查看: 2449 | 回復(fù): 20 | ||
| 當(dāng)前只顯示滿足指定條件的回帖，點(diǎn)擊這里查看本話題的所有回帖 | ||
東樂樂金蟲 (初入文壇)
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[求助]
用組織提取總RNA后逆轉(zhuǎn)錄得到的cDNA進(jìn)行PCR一直未有目的條帶，請(qǐng)大家?guī)兔Γ? 已有3人參與
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我打算做質(zhì)粒構(gòu)建，但是第一步PCR就進(jìn)行不下去了。采集新鮮組織后立即用Trizol法提取RNA，用takara的逆轉(zhuǎn)錄試劑盒得到的cDNA，用師兄師姐留下的引物進(jìn)行PCR可以擴(kuò)增出相應(yīng)大小的目的條帶（約1500bp），但用我自己設(shè)計(jì)的幾對(duì)引物都不能擴(kuò)增出條帶（目的條帶大小：3564bp），只有最下面有疑似引物二聚體的條帶，其余的一條帶也沒有。我想要獲得該基因的CDS全長(zhǎng)，之后進(jìn)行細(xì)胞轉(zhuǎn)染。選的酶切位點(diǎn)和載體都是實(shí)驗(yàn)室里現(xiàn)有的。實(shí)驗(yàn)新手有很多不懂的，請(qǐng)大家?guī)臀蚁胂朕k法，謝謝大家！ 克隆載體：19T【T-Vector pMD19(Simple)】 表達(dá)載體：pEGFP-N1 引物：（其它引物跟這個(gè)差不多就是上下幾個(gè)堿基的增減） 上游：5‘-CCCAAGCTTGCCACCATGCCTTTTGGTCTGAAGCTC-3‘[Hind III酶切位點(diǎn)，我引入了Kozak序列(GCCACC序列)，據(jù)說這樣有利于以后的表達(dá)] 下游：5’-CCGGAATTCTTAAATGAGTCTGGAGTTTTGGAGG-3’(EcoR I酶切位點(diǎn)) 體系：共25μl 高壓過的純水 16μl 10X LA PCR Buffer II (Mg2+ plus) 2.5μl dNTP Mix 2μl 引物上游、下游各1μl cDNA模板 2μl Takara LA Taq 0.5μl 條件：35個(gè)循環(huán) 94℃ 2min（3min也做過） 94℃ 30s 55℃（58℃和61℃也做過）1min 72℃ 4min 72℃ 10min 4℃ ∞ CDS序列：共3564bp ATGCCTTTTGGTCTGAAGCTCCGCCGGACACGGCGCTACAACGTCCTGAGCAAGAACTGCTTTGTCACACGGATTCGCCTGCTGGACAGCAATGTTATCGAGTGCACGCTGTCGGTGGAAAGCACAGGGCAAGAATGCCTGGAGGCTGTGGCCCAGAGGCTGGAGCTGCGAGAGACGCACTACTTTGGCCTTTGGTTTCTCAGCAAGAGCCAGCAAGCACGATGGGTGGAGCTGGAGAAACCTCTGAAGAAACATCTGGACAAATTCGCTAATGAGCCTTTGCTTTTCTTTGGAGTCATGTTCTATGTGCCAAATGTGTCATGGCTTCAGCAAGAGGCCACAAGATATCAGTATTACCTGCAAGTCAAAAAAGATGTGCTTGAAGGGCGATTACGATGTACATTGGACCAGGTGATTCGGCTAGCCGGCCTAGCTGTGCAAGCTGATTTTGGAGACTATAATCAGTTTGATTCTCAAGATTTCCTCAGAGAGTATGTGCTATTTCCTATGGATTTGGCCCTGGAAGAGGCTGTTCTGGAGGAGCTGACCCAGAAGGTAGCCCAAGAACACAAAGCCCACAGTGGAATCCTGCCAGCAGAAGCTGAACTGATGTACATCAATGAAGTTGAACGTTTGGATGGATTTGGACAGGAAATCTTCCCTGTAAAGGACAATCATGGAAACTGTGTACACCTTGGCATTTTCTTTATGGGGATTTTCGTGAGGAACAGAATTGGAAGACAAGCGGTAATATACAGGTGGAATGACATGGGGAATATCACTCATAACAAGTCGACCATTCTAGTGGAGCTCATCAACAAAGAAGAGACTGCCCTCTTTCACACGGATGATATCGAAAATGCCAAGTATATTTCTCGGTTGTTTGCCACACGACACAAGTTTTACAAACAAAACAAAATCTGCACTGAACAGTCAAATTCTCCACCCCCCATCAGACGCCAGCCCACCTGGAGCCGATCCTCTCTGCCCAGGCAGCAGCCGTACATCCTGCCTCCCGTTCACGTCCAGTGTGGTGAGCACTACTCGGAAACGCACACCTCGCAAGACAGCATTTTTCATGGGAATGAAGAAGCCTTGTATTGCAACTCTCACAACAGCCTGGACTTAAATTATTTAAATGGCACTGTCACCAATGGCAGCGTGTGTAGCGTTCACAGCGTCAACTCCCTCAACTGCTCGCAAAGTTTCATCCAGGCCTCCCCTGTATCCTCCAACCTCAGTATCCCTGGGAGTGACATCATGCGGGCCGACTACATCCCGAGCCACCGGCACAGCGCGATCATCGTGCCCTCGTACAGGCCAACCCCCGATTATGAGACAGTCATGCGCCAGATGAAGAGGGGGATCCTGCATACAGACAGCCAGAGCCAGTCTCTGAGAAACCTCAACATTATCAACACCCATGCCTACAACCAGCCAGAGGATCTGGTGTACAGCCAACCGGAGATGCGGGAGAGGCACCCCTACACTGTCCCTTATGGGCCACAGGGGGTCTACAGCAACAAACTTGTCAGTCCATCTGACCAGAGGAACCCAAAGAATAATGTGGTACCAAGCAAGCCGGGGGCAAGCGCCATCTCGCACACGGTGAGCACCCCAGAGCTGGCCAACATGCAGCTGCAGGGCAGCCATAACTACAGCACGGCCCACATGCTTAAGAACTATCTCTTCAGGCCACCGCCCCCCTACCCACGGCCACGACCTGCCACCAGCACCCCAGACCTGGCCAGCCACCGCCACAAGTACGTCAGCGGCAGCAGCCCGGACCTGGTGACCCGGAAGGTGCAGCTCTCGGTGAAGACCTTCCAAGAGGACAGCTCTCCGGTGGTTCATCAGTCTCTCCAGGAGGTGAGCGAGCCCCTCACGGCCACCAAGCACCACGGCACTGTGAACAAGCGCCACAGCCTGGAGGTGATGAACAGCATGGTGCGGGGCATGGAGGCCATGACGCTCAAGTCGCTCCACCTCCCCATGGCTCGCCGCAACACGCTCCGGGAGCAGGGACCGCCCGAGGAGGGGTCAGGCAGCCACGAGGTCCCCCAGCTCCCTCAGTATCACCACAAGAAGACCTTCTCTGATGCCACTATGCTAATCCACAGCAGCGAGAGTGAGGAGGAGGAGGAGGAGGCTCCAGAATCGGTGCCCCAGATCCCCATGCTCCGGGAGAAGATGGAGTACAGTGCCCAGCTGCAGGCGGCCCTGGCCCGCATCCCCAACAAGCCCCCGCCTGAGTACCCCGGTCCAAGGAAGAGTGTGAGCAATGGGGCTCTGAGGCAGGACCAAGCCAGCCTTCCTCCCGCCATGGCCAGAGCCAGGGTGCTGAGGCATGGGCCAGCCAAGGCCATCAGCATGTCTCGGACTGACCCGCCGGCTGTCAACGGGGCCTCTCTCGGCCCATCCATCTCGGAACCCGACCTGACTAGTGTGAAGGAGCGGGTCAAAAAAGAGCCTGTGAAGGAGAGACCTGTGTCTGAAATGTTTTCCCTGGAAGACAGCATTATAGAGAGAGAGATGATGATCAGGAATCTAGAGAAGCAGAAGATGGCAGGCCTGGAGGCACAGAAGAGGCCGCTGATGTTGGCAGCATTGAATGGGCTCTCGGTGGCTCGAGTCTCAGGGCGGGAAGAGAATCGAGTTGATGCCACCCGGGTTCCCATGGACGAGAGGTTCAGAACCCTGAAGAAGAAACTAGAAGAGGGAATGGTGTTCACAGAATATGAGCAAATTCCAAAGAAAAAGGCGAATGGCATTTTCAGCACAGCAGCTCTGCCAGAAAACGCCGAGCGCAGCCGAATCCGTGAAGTTGTCCCCTATGAGGAGAATCGAGTAGAGCTGATACCAACCAAAGAAAATAACACAGGATACATTAATGCCTCCCACATCAAGGTGGTGGTTGGCGGGGCAGAATGGCACTACATAGCCACCCAGGGGCCCCTGCCACACACGTGCCACGACTTCTGGCAGATGGTGTGGGAGCAGGGAGTGAATGTGATTGCCATGGTCACTGCAGAGGAGGAGGGTGGACGAACCAAAAGCCACCGATACTGGCCCAAACTAGGTTCAAAGCACAGCTCAGCCACCTATGGCAAGTTCAAGGTCACCACGAAGTTTCGAACGGATTCTGTTTGCTATGCAACCACGGGCTTGAAGGTCAAGCACCTTTTGTCTGGGCAAGAAAGGACGGTGTGGCATTTACAATATACTGACTGGCCAGATCACGGCTGTCCAGAAGATGTCCAAGGATTTTTATCCTACTTGGAGGAGATCCAGTCGGTCCGTCGCCATACCAACAGCATGCTGGAAGGCACCAAGAACCGGCACCCGCCCATCGTGGTCCACTGTAGTGCTGGGGTGGGAAGGACCGGCGTGCTCATTCTTTCTGAGCTGATGATCTACTGCTTGGAACATAACGAAAAGGTGGAAGTGCCCATGATGCTGAGGCTCCTCAGGGAGCAGAGGATGTTCATGATCCAGACTATCGCTCAGTACAAGTTTGTCTACCAAGTCCTCATCCAGTTCCTCCAAAACTCCAGACTCATTTAA 總的mRNA序列：其中654bp至4217bp為CDS區(qū)域 ACTCGCACGGCCCCTTCCTCCCTCCTCTCCCGGCCGCTCGCATTTCCTGCCGCTCTGGCTCTCCCGGCCCCTCAAAGTTCTTTCCAACTTTTTCTCGGCGGAGTGAGCGCAGCGGGCGCAGACTCGGGGGCAGGTTGCTGTGCTTCTCCGGGCTCAGCCGCCTGCTCTCCTGGCTCAGGTCCTCGGGGAGCCCTAGACAGACATCAAGTGGCCACTGGCGCTCCTTCCCCTCCCAGCTGAGCCATCCTCCCCGGCCTCCTCGGGCGGGACAGCCCCGTGCTTAGGTTTTTCTCCTTTTCTCCCCCGGTGCGCCTCTGCTCGGACTCTCGCGCCGGGATCGCGGCGGAAACCTCCCTCCCCTTTCGCCTCCTGCGGCTCCTTCCCTTCGCCCCTCCTCCGCCAGTCACTGGAATCAATTCCGTGGGGAATCGGCTCCGCCGCCGCGAAGGACAGCCTTTCCGCGCGGGACTCCGGGGCGCCACGGGGGCCATGTAAGCAGCTATCTTCCAGAGGGCCACACTGGGCATGGACACCCTTTTCCCTGCCTGGAGGAGCACAGGTGATAGTGTAATTTTCCAGTCACGAAACTGCTAAGGCCATCTCAGGGGCGTGTGCGCCAGGATAGGCGGGCGGCGTCCGAGGACCACATAGCCATGCCTTTTGGTCTGAAGCTCCGCCGGACACGGCGCTACAACGTCCTGAGCAAGAACTGCTTTGTCACACGGATTCGCCTGCTGGACAGCAATGTTATCGAGTGCACGCTGTCGGTGGAAAGCACAGGGCAAGAATGCCTGGAGGCTGTGGCCCAGAGGCTGGAGCTGCGAGAGACGCACTACTTTGGCCTTTGGTTTCTCAGCAAGAGCCAGCAAGCACGATGGGTGGAGCTGGAGAAACCTCTGAAGAAACATCTGGACAAATTCGCTAATGAGCCTTTGCTTTTCTTTGGAGTCATGTTCTATGTGCCAAATGTGTCATGGCTTCAGCAAGAGGCCACAAGATATCAGTATTACCTGCAAGTCAAAAAAGATGTGCTTGAAGGGCGATTACGATGTACATTGGACCAGGTGATTCGGCTAGCCGGCCTAGCTGTGCAAGCTGATTTTGGAGACTATAATCAGTTTGATTCTCAAGATTTCCTCAGAGAGTATGTGCTATTTCCTATGGATTTGGCCCTGGAAGAGGCTGTTCTGGAGGAGCTGACCCAGAAGGTAGCCCAAGAACACAAAGCCCACAGTGGAATCCTGCCAGCAGAAGCTGAACTGATGTACATCAATGAAGTTGAACGTTTGGATGGATTTGGACAGGAAATCTTCCCTGTAAAGGACAATCATGGAAACTGTGTACACCTTGGCATTTTCTTTATGGGGATTTTCGTGAGGAACAGAATTGGAAGACAAGCGGTAATATACAGGTGGAATGACATGGGGAATATCACTCATAACAAGTCGACCATTCTAGTGGAGCTCATCAACAAAGAAGAGACTGCCCTCTTTCACACGGATGATATCGAAAATGCCAAGTATATTTCTCGGTTGTTTGCCACACGACACAAGTTTTACAAACAAAACAAAATCTGCACTGAACAGTCAAATTCTCCACCCCCCATCAGACGCCAGCCCACCTGGAGCCGATCCTCTCTGCCCAGGCAGCAGCCGTACATCCTGCCTCCCGTTCACGTCCAGTGTGGTGAGCACTACTCGGAAACGCACACCTCGCAAGACAGCATTTTTCATGGGAATGAAGAAGCCTTGTATTGCAACTCTCACAACAGCCTGGACTTAAATTATTTAAATGGCACTGTCACCAATGGCAGCGTGTGTAGCGTTCACAGCGTCAACTCCCTCAACTGCTCGCAAAGTTTCATCCAGGCCTCCCCTGTATCCTCCAACCTCAGTATCCCTGGGAGTGACATCATGCGGGCCGACTACATCCCGAGCCACCGGCACAGCGCGATCATCGTGCCCTCGTACAGGCCAACCCCCGATTATGAGACAGTCATGCGCCAGATGAAGAGGGGGATCCTGCATACAGACAGCCAGAGCCAGTCTCTGAGAAACCTCAACATTATCAACACCCATGCCTACAACCAGCCAGAGGATCTGGTGTACAGCCAACCGGAGATGCGGGAGAGGCACCCCTACACTGTCCCTTATGGGCCACAGGGGGTCTACAGCAACAAACTTGTCAGTCCATCTGACCAGAGGAACCCAAAGAATAATGTGGTACCAAGCAAGCCGGGGGCAAGCGCCATCTCGCACACGGTGAGCACCCCAGAGCTGGCCAACATGCAGCTGCAGGGCAGCCATAACTACAGCACGGCCCACATGCTTAAGAACTATCTCTTCAGGCCACCGCCCCCCTACCCACGGCCACGACCTGCCACCAGCACCCCAGACCTGGCCAGCCACCGCCACAAGTACGTCAGCGGCAGCAGCCCGGACCTGGTGACCCGGAAGGTGCAGCTCTCGGTGAAGACCTTCCAAGAGGACAGCTCTCCGGTGGTTCATCAGTCTCTCCAGGAGGTGAGCGAGCCCCTCACGGCCACCAAGCACCACGGCACTGTGAACAAGCGCCACAGCCTGGAGGTGATGAACAGCATGGTGCGGGGCATGGAGGCCATGACGCTCAAGTCGCTCCACCTCCCCATGGCTCGCCGCAACACGCTCCGGGAGCAGGGACCGCCCGAGGAGGGGTCAGGCAGCCACGAGGTCCCCCAGCTCCCTCAGTATCACCACAAGAAGACCTTCTCTGATGCCACTATGCTAATCCACAGCAGCGAGAGTGAGGAGGAGGAGGAGGAGGCTCCAGAATCGGTGCCCCAGATCCCCATGCTCCGGGAGAAGATGGAGTACAGTGCCCAGCTGCAGGCGGCCCTGGCCCGCATCCCCAACAAGCCCCCGCCTGAGTACCCCGGTCCAAGGAAGAGTGTGAGCAATGGGGCTCTGAGGCAGGACCAAGCCAGCCTTCCTCCCGCCATGGCCAGAGCCAGGGTGCTGAGGCATGGGCCAGCCAAGGCCATCAGCATGTCTCGGACTGACCCGCCGGCTGTCAACGGGGCCTCTCTCGGCCCATCCATCTCGGAACCCGACCTGACTAGTGTGAAGGAGCGGGTCAAAAAAGAGCCTGTGAAGGAGAGACCTGTGTCTGAAATGTTTTCCCTGGAAGACAGCATTATAGAGAGAGAGATGATGATCAGGAATCTAGAGAAGCAGAAGATGGCAGGCCTGGAGGCACAGAAGAGGCCGCTGATGTTGGCAGCATTGAATGGGCTCTCGGTGGCTCGAGTCTCAGGGCGGGAAGAGAATCGAGTTGATGCCACCCGGGTTCCCATGGACGAGAGGTTCAGAACCCTGAAGAAGAAACTAGAAGAGGGAATGGTGTTCACAGAATATGAGCAAATTCCAAAGAAAAAGGCGAATGGCATTTTCAGCACAGCAGCTCTGCCAGAAAACGCCGAGCGCAGCCGAATCCGTGAAGTTGTCCCCTATGAGGAGAATCGAGTAGAGCTGATACCAACCAAAGAAAATAACACAGGATACATTAATGCCTCCCACATCAAGGTGGTGGTTGGCGGGGCAGAATGGCACTACATAGCCACCCAGGGGCCCCTGCCACACACGTGCCACGACTTCTGGCAGATGGTGTGGGAGCAGGGAGTGAATGTGATTGCCATGGTCACTGCAGAGGAGGAGGGTGGACGAACCAAAAGCCACCGATACTGGCCCAAACTAGGTTCAAAGCACAGCTCAGCCACCTATGGCAAGTTCAAGGTCACCACGAAGTTTCGAACGGATTCTGTTTGCTATGCAACCACGGGCTTGAAGGTCAAGCACCTTTTGTCTGGGCAAGAAAGGACGGTGTGGCATTTACAATATACTGACTGGCCAGATCACGGCTGTCCAGAAGATGTCCAAGGATTTTTATCCTACTTGGAGGAGATCCAGTCGGTCCGTCGCCATACCAACAGCATGCTGGAAGGCACCAAGAACCGGCACCCGCCCATCGTGGTCCACTGTAGTGCTGGGGTGGGAAGGACCGGCGTGCTCATTCTTTCTGAGCTGATGATCTACTGCTTGGAACATAACGAAAAGGTGGAAGTGCCCATGATGCTGAGGCTCCTCAGGGAGCAGAGGATGTTCATGATCCAGACTATCGCTCAGTACAAGTTTGTCTACCAAGTCCTCATCCAGTTCCTCCAAAACTCCAGACTCATTTAATCACCCCAATCCAGCTCCTGGAGGAGGGACCCAGCTCCATCGCGCTGGAGGAGAGTCACCTCCAGACAACATCTGCTCCCCCCACAGGGGTGCAGGTGGCTGGCAGCAAACAGGCTCTCTGAAGACAGTAGCCAAGATTATTCACACATACCATGTATTATTTTATATGAGATAATTTATTTTTTTCCCCTTTGGAATAACTTTTGTGAATTATTATAATGCAGTTTCCCTAGTAATATAGTACTTTTCATTTGAACCACATCTTGACTGATCTGTATTGTAATATATGTCAGCAGGTAAGGTTGCCTGCTGGATCATTTTGAGGACAGAGGCATGAGGGAGCACATCTCTTGTGAAGTTGCAGCCAGATTTGTAACCAACCCTGAAATTCATCAGCTTAATTCATTTATCAGCTTGATTCATTCATCATTCATTGCTTATATCCAAAGCAAAGACGGTAAGAAAATGAATTCATCCTGAAATATAAAGAAAAGGGTCTGAAGGAACAAACACGATTCTCTTATATTTTGGGGCTCATGAGCCTTGATAGACAGTTTCCTCTCGTCTTCATTTCCACCCCTCATCCTCAGTAGTCTCCTCTCCCCCACGCCCCACCCCAACTTCCCCCCCAAGCTTGAGTTAAAGACAGAATAGCTAAAGACAGTGCTGCCTTTACAATGCAGTAATTGCCATCTTTGGGGCCGAAAGACAAGCTCTGTGTTGTGCTTTTCTTGACCACCCCTTATCCTGGGCTCTGGAGCTTGTGTTTCCCTGCTGGCGACTGTACCTTGGGTATTTGTTGCTACCTCTCCTGTTTGCTCAGTAGGACCCTGTCTGGTGGCATTGAGGCTCTGGACCAGACCATCTGTGCAGTTAAGGCTCTACCCTGATTGAGAGAGGATAGCAGACCTAGAAAGAGAAAGGAGTTGGGCAGGGCCTTTGAGGATTGTGTTTTTCAGGCAGGGCCTTGATGATCATTGTTTTTTATTTAAATAAGATGTGTGTGCTGGACAGAGACCTAAAAGTTGAGGTCACTAAGTCATTGGAAAGGCCATCAAGGAAACAGATGGGGAAGCTGATTTATGGGAGCTGTAAGGCATTTAGCTACATAACAGGGGTCCTGGCCAGGAAACACATCAAATGTGACCCCCGCTGTGCTGATATCATCTTCAGGCTTTGGTCTGCAAGATCAGAATTAATCCCACTCGGGACCCCATAGTCCAAACTTGGGGCCACTTGATGAACGATGGTAGAATTGTCATTGGCAGAGCCCTGTGCTTCTTTCCTTTTCTTCATAAAATCCACTCGCTGGTCAGTTATCTTCACTTTGAAGCCCAGTTCTTAGTTTCTTCCTATGGCTTCATTGGTCAGTGTCCTTCTGAATTTCCAAGGATGGTACACAATAAATCATGTTTTGTACTTTTTTCCTCTTACTGCATTTTGGGGGATTTATCATTCTATGTCTACCTTTTCTTGAGTACAGCTTTGATATGCACCTGTTGTTACGTGGTGATGGGAAGTCACAGGCGTGCTCTTTCTAGTTAATTTGATGCCACATCTTCCTTGTCTTTTCAGCTTGGGAAAAAGGCGGCAGTGGAGGAAGGCATGGAATGCCCACAGTGGTCAGTTCAAAGAACAAACGTGCAATTAAAAAACTGTAGTCAGCCAGGCACGGTGGTTCACACCTGTAATCCCAGCACTTTGAAAGGCCAAGGCGGGCAGATTGCTTGAGCTAAGGAGTTCGAGACCAGCCTGAGCAACATGATGAAACCCCGTCTCTACAAAAAGTACAAAAATTAGCCAGGCGTGGTGGTATGCCCTGGTAGTCCCAGCTGCTCGGGAGGCTGAGGCAGGAGGATCACCTGATTCTAAGAATTCGAGACTGCAGTGAGCCGTGATCTTGCCACTGTAGTCCAGGCTGGGCTACGGAGAGACCCTGCCTCCAAAAAAAAAAAAAAAAAAGGAAAAAAGGTTGTCAAGAAAAACTAGATGTTAGGAGAAAGGAAAATTTAATTGCAGTTTTTTTTCTTAGAATTGACTGCTGTGAGAGTTCCATATGCCTTTCTTCATTGCTGCTTTTGTCCCCCGTGAGCTAAAAAGATGGAGTGACATCAAATCAACCAGAAAAAGTATGCCTTTGTGACATCCCATCACCACATGCCAACAGGTATATATTCCCCATTAAGTTCTTCGGAATAGGAATCCTCTGTTTCAACCTGGCCAGGTGTTGTGGTGGCTGTACTCTAGTTAGACTCGGAATATCTGGGGATGGAGGGCTTCCCCTGTGTCTTCTACTTCAAGGTCTGAAGGCTCAGTGAAGGAGTATAATCTGCTGATCTTTGTAGATTCTGGAGTTTTGTTGTATGTCCTGGAAAGAAACCCATTAGTATTACATGTATTTTCAGTGAACAGAGCTTATAACCCTTATTATAAGAAGCTCATCAATAAGCAAAAAGATACTTGTTTCCTTTCCTTGGAGGTTTTTCCATCCTTGGGATATTCTGCTGTTAGGGATGTTTTAGCAAGTGGTCTCAGTTACTGGTTTATTGCGTGATGAACAACATCAGTATTTATCTTTTATCTCTAAGCCCCAAGGTGGGCACTGTTAGAATATGTCTCATGTGGACAGCATATAGATCTGGTGCGTCTTTGAGGTCGTCAGAGCTCATGGGCTTCCCTGAAATTCATCCACTGTCCCTGCCGTATGCTACGGGAATATTCATTAGTGTACAAAATGCAGGGAGGAAGTAGGTTTAATATTCAACTTTCTAGCCAAAGTTTATATTGAAACCCAAAAGAAAACATTTAAGAGTTGTTCCACATATTTCACTTTTAAAAACAAATGCCTTTGGTTCTTTAGCACATTTTGCATTCCTTTTCACATCTCCAGTAAATGCCAACATATCTCCTGTTAAATTAGCAGCAGCCATTTAAAGTCCTTTCGGTGGCATCTGCATAATAATTGCCCAGAGATGCTTTATATCTGGGAAGCAAGCCAAGGAATAAACCTTGAAGCAAAGTGTATTAAATTAGTTATCTAGTTAGAGCTTTTGGAATGATTTCCTGATGATGTATCAAGTCTGAAGCTGGAGCTGTCAGTGTCTATTGCTGCAGTTTGGATTTGAAGGGAGAAAATGTAAAATGGAGGAAAAAAAAGTTACCATCTCACAACAAAGCCATCAAACATTTTCCAGCCGCTGTTTTCGAGGTTTTCCAGTTGAACTGTTTGGTTTCTTTCATCCACACTCATTTGGATACATTGACCCGAGGTATTCATCCTTGTTTACTGTGGTCCCTGAATCATGGGGGCTGAATTTGATGTCTTCATCCTTGAGATGAGCCTGCTGGCTTAGCTGAGGAATGTCCTGCTGAGGTTTCTTAGGTTTCCTTGGGTTCTAAGGATATACTGGATATACCATCTTTTAGCAAGAGTATCTGGTAGCATTTACAGATAGCATAGACATTGGTATGCACTTCTTTCCCCAGATAGGAAGTAAAGGAGGATTTAGTTGCATGAAAAAAGGATGTTAAACATTGATTACATAGGAGTAAAGATGAATGAGCTGCAATATTCAGTCGGAGCTAAACAATAAGATCAGGGAAGGTAAAAATACCTATGTGGAATATTTTGAATCGTAAGCTTTTGAGGAGCTTAAATTGAGAGAATTTTACTTTTAATTTTGTAGATTGAGAAGAGGAACCGCTTTTTAAAATTATAGCTAAACTGTCATTGTTTTCCTAAGAGTCACTTGGCCATCTCTGGCCCCCTCTTTCATCAGCCTGAAGAGAGGGTCTTTGTAGACTGCTGAGGGTGGGCCTTGTAGGACTTGACCATGGCTTACACCTACTTAACCTTTATCCTGCTTTCTTTCAGCTTGTGCTTTTCAGTTATAAACTCCAGTGGGTACAGCAGGCTGGCCTTTTCATCCAGCTGATTATTTTTCCAGCTTAATATAGATTGACCCATATGAAATTTCCAATAATGGACCATATTTTCTGCAAATAGACAGTACTCGCATGGATCACCTATATCTTCCCCCTGATACACTGTGGGTCCCAACACCAGATGTCATTTCTCCAGAGCAGTGCTAATGAACACAAAAGGTATACCCTGGGTGGCCCAGCTCTTTTCACGAACGTGCTGCCCTGCTCATAGTGATCCTTGCATCACTTGGTGAATGGGCCATCTCCTGGGACATGGAAGTTGCAGAGGTAGATAGTGCACTGCAGCTTCTCTTAAGCCGGATTGGCCATCAGGCATATCACTCTGGAGTTTTTAGCTGCTGTCCTTTCCCGATGAACAGTCTGTATTAGCTGACCTCAGCCTACTTGTTACGTGACGTATGGGTCCCAAAAGTGTCCTTTGTCAAAAAGCAGAATGTGCCTTCTAGTCTCCCTTTCCCCATCTAATGGTGTATTCGATGGTGAAGATGAGTACAGTTGACCACCCCTATCTGTGGGTTCCACATCCCTATATTCAACCAAGTACAGATTGAAAAATATTTGGGGAAGAGGGGAAACCCACAAAGTTCCAAAAAGCAAAAGTTGAATTTGCCACATGCTGAATACTACATTGAATTCACACAAATGAAGTGATGTGTAGGCATTGAGTTAGGTATTGTAAATAATCTAGAGATGATTTAAAGTATGCAAGAGATGTGCATAGGTTATATGCAAATACTGTGCTATTTTATATAAAAGACTTGAACATCCATGGATTCTGGTATTCTCAGAGGGTCCTTGATTGCCCCCTTTGGTAAAGGACAACTATTTCGTTACTGATTTTCGTTTGGGGAAGATCTGTCAATCCCTTGAGGTGCGGGGGTTGGGGGGATGGAGGGTACAGGGCATTCTAGGATGTGTGCCAGGGAGCACAGATTCAAGGGATGGGATTGAGTCAGACCTGTGTTCTTACTCAGTGTCAGAAATAACTCTGTGGAGCTCCTAGAGTAAGACATTTCCGGAAGCACCACATAATTACTGTTGGGCTCTTAGGGTAGCCCTTTTAGGGAATTGAGCATTCCCATGTTTTACCAACAATTATTCTGCTGCTGTGTTTTATTATATTGCCAATGGTTTTGAGACACTCATCATGCTCTTATTTAGTGATTTCTTTTCATGAGCAGAGCAACAGCTCATCCAGCATGGTTTCCAAATGGAGAAATTTGGGTCTTCTGTAGAAACCACACAAATTCTCCAATGGCCTACAGCCTTATGGTTGGCACACTAGTTGGCCCTATAGGGTGGAAATAAAGCTGTAAGATGTTAAACTGCATTTGATACTTCTCTTGAACGCTGAGCAAGGAAGCAAAATAGTTCTTGTCTTTACTTAAGCTTCTAAGACATTTTGGGGCAAAGGACCTTACAGATGGCGTCTGTTGAAAGTAACAGCAACGTGCCAGGGAGAAATGTGGGGGAAATCTCATCAAATTCTGCCACCTCAAATGTGTTGTCCAGAAGTCAGTGTTATTCAGGGGGCCCTGTGAAACTTGACCACCAGCTTGCGCCATCAACACTCAGCCTTTATCCAGCCTGCTCTCAGCTTTTGTGTTTCTGTTAGAAACTCAGATAGGTAAGTATTTTTATTCAGTAGACTACCTTTCTTATCTTTTCAGCTTAATATAGCTGCATCTTCTTTCTCAAAGCCAAACTAAGATATTCTCTATTAAAATGTCCATGAGCCTAGCATTGAGTGTCTGGCATCCATCAATTTCATAGACTGGAAAATGATTGTTGTTTGGTACAGTAAAGAAGAGGATGTGCATCAGTTCCTACCGTTTGCAGCTTTGTTTTTAGCCTTCCCATTTTAAAAATAAATTCAAGGACATTGAGTTACAAGGCAGGAGGGCTGGAGCTACTGGGCAGCCTGAATATGACAAGCATTTGTGTGGAAAGTCATTGCTCCTTCTGCCACACTTTGGGCCATCAGGATCATTCTTTCCCAGAAGTGCCATAAACTTGCTCAAAAGTTTCTATAAATGGGAAGAGGGAGAGGAAGGATTTTTGCATCAGTCCTGAAGTTGCTATCCAAAAGTTCTCTGTTTCACAAATAATTTTCTGAACTCTGGAATGCCTCTCCTACTCCCTGCCTCCCTTTCTGTAATGTCAGAGTGATGGAAACCACCAAGTGGCATGCTAGGGAAAGCCTGCAGCAGTGTTGGAGTCTATTTCACCCTAGCTCATAGTTTTAAACTGTCTTCACTGTTGAGGTAGAGCTTGATGAATGTCATGGATTATGATGTGTGGTTTATCATATTTGCCTGGATTTGCTGATCAAAAGCACCATCTTCCCTCGCCTGCTGCTGGCAGCCTTTCCTTGCCTTGCTTGTTAGCAGAGCATTCTGCTTACCCATGTGGCTCCCAGAGTTAGCAGCCCCGGCTCTTGGATTTCTTGATTCTTCTCCCCTGTGATCTCAGAGGTGCTGCAGAGGACATTCCCCTTTAGAGCAAGTCATGTTTCTATTCAGGCCACAAAACTGGGATGTACATGCAGTGACTTTGGTGTTCCTTGTCTTGTTCAGGGGAACGGGTGGGACTGTTGTGTGCTGTCACCCTCTTCATTCCATGAGCACCTTGTTCACTTAGGGTCTGCTGCCTTTTTTTTTTTCTTTTTCTTTTTTTTTTTTTTTTTAGTTTTTGAGATCGTGTCTCACTCCTGTTGCGCAGGCTGGAGTGCAGCCTCCAAAGTAGCTGGGATTATAGGCGTGCACCACCACACCCGGCTAATTTTTATATTTTTAGTAGAGACGGGGTTTTGCCATGTTGGCCAGACTGGTCTCAAACCTCTGACCTCGTGATGACCCCCCTCGGCCTCCCAAAGTGCTGGGATTACACCCAGCCTCTGCTGCCTTTTCATATTTCCCCATCTGCTTTATGGATCAACTCTCAACAGTACACTTTTTCTTTTCTTTACCTACCCTATGAGTGCAACCCAGATGTAAGAGTTAATCCTCGTCAGAGAATCATTGCCTTAAACCTCTCAGAAATATGTAATTAGGAAATCTTATTTTAATTTTTTAAAAATTGCTTGTATAGTTTCAAAGAATAAGATCTGGCAAATGGCCAGATGTGGTGGCTCACACCTGTAATCAAACCAGCACTTTGGGAGGCTGAGGCAGGCAGATCACCTGAGGTCAGGCGTTCAAGACCAGCCTGGCCAACATGGTGAAATCCCGTCTCTACTAAAAATACAAAAAAAAAAAAAAAAAAAAAAATTAGCCAGGCATGGTGGCACATGCCTGTAATCCCAGCTACTCAGGAGGCTGAGGCAGGATAATTGCTTGAACCCGGGAGGTGGAGGTTGCAGTGAGCCGAGATCATGCCACTGCACTCCAGCCTGGGCAACAGAGTGAGACTTTGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAGATCTGGCGGATGAAAATAACCAGAATGAAAATAGCTAGAAAACTCAGCAAGCAGGAAGCTCCCTTTCTCACCCTTTTGTTCCCTTGCCGATAGAATCAGTCACTATTAGAAAAAATGAAAGACGCTCTGTTTAAAACAATGATGACAGCAGTACTTAATATGTATTTCGAGGTGAACTTATATAGATTGAGAGAGGCTGCATTTGGCAGACTGATGTATAGGAAGACCCATTTGTTTCTAGCTTCTCCCTGCAGGGAAAATGCTTTCGTCATTATAGCCTCTTTACACAGACTGGCCATTCTAGTGAAACAGGTGGTAAACCTTTGGGCTGCCCAGAAACATTTTATCTGTTTTCACTTACCTAGGAAGGGGAAAGATTAGCGGGTCATCCAAAATCTGTATGTAAGCTATCTTCATTTTCTTCCCCAACCTTCTCCTCCTGGGAAACACAAATGCTATCTCATCTGACAAAAGGTTTTAGAGGATAAAGCTGAAAAGATTGGATTGGGATCTTTTTGTGGCTTGGGGCGGAGCCTTTTGCTAAAATCTCAAGAATGCTGCTTTGAGTTTAGCTAGGGTGGCTCTCAGAACTGGGGTGCCTGGCATTCTCAGCATTTCTCAGGGGCCTCCCACCTCTGACAACTGCAGTGTTAGCTAATACATACCTTGAGCATAGAACTGAATGCTGTAATTCAGAGCCATTTTTTTTTTCAACTTGAACATTGTACAATTTTACTGCAATTTCCTTTGAACTTTCTTGCCACTGTTTGGAATCTTAAAAATTCATTAGCCTTCTCCTTTCTGACATAAAGCTACTCTTCATCAGAGATGAGTTCCTATGTATGTCCTTTGTTCCTTCAATAGCTAATTAATGTGCTTGAGGATACTTCAGTGGAAAAAAAGGTTTAAATATGCAAATTACTAATAAATGTGTAACCTTATGTAACTTGTGTTACATCAAGTAACAAGCTAATCTAGTTTGTTTCACTGGACTAGGCTTGTGCTCCCTACTTCAGTATTTTGATGCTTTCCTTGATCTTTGTTTCACAAAATGTTGTGAATTTTGGTATCATTCAAAACAAATGACATTTATTAGGTTTCATTTTGAAACGATGTACAGACAAGTCCCCAACTTAGAAACCGGTTTGTTCTTAAGGTTCTTGCGTCAGCCCATAGAAGCCCACTGACCTCCACCACAGCCCAAATGGAGGGCTGTGATAGCCAGATCTGGTTGGCTTTTGTGGGCTGACCCAGACATTTAATCACCATCTCTTATGTTGTTGCCGTAAGAAATGCATTCCAGGTTGGGACTTGGGATCCTGAGAGCACATTCGCCCCCTGTGGTGGCCGCTTGCCACCTTGCAAGATGGAAGCCCAGTCTCCTTACTACCAAACTGTAGTTGTAAGCAGAGGGAGGGGTGAGATGTTTATAGGACATTCCCTAAGCTGGGGAGTGATTTTTATCACTATTCATGTCAACTGTACTTTGGTATAGACTCCCTATCAATTTAATAATATGAAAAGCCTAAAATAAAACTATGCATGCTATTCTATGTGCTATTTTATATCAGTAAATAAGCTTATGCTTGCCAGTTGTATACACAGTTATGAGGTGTATAGAACTGACTTTGACAGTATTTTTTGCACTGTTTCCTATCTGTTTTTATAAAGTCTTATTTAGATATTGGACCTTGTTGATGTTCTCACTGCCCTTGTGCTTGCTATAAAATGTTTCATATGTGCCTTTACAAATGTGAGATCTTTATTCTAACCTTTTTTTGTAAAAGATATCTATTGATTTCCATATGCAATAAACCTTTTTTTCAGAGAAAAGTTA 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東樂樂5樓2015-10-19 12:07:31
liping121200
銅蟲 (小有名氣)
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- 金幣: 129.7
- 紅花: 1
- 帖子: 82
- 在線: 39.3小時(shí)
- 蟲號(hào): 4060168
- 注冊(cè): 2015-09-09
- 性別: MM
- 專業(yè): 生物大分子結(jié)構(gòu)與功能
2樓2015-10-16 15:45:27
東樂樂
金蟲 (初入文壇)
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- 帖子: 50
- 在線: 29.5小時(shí)
- 蟲號(hào): 3562569
- 注冊(cè): 2014-11-27
- 專業(yè): 流行病學(xué)方法與衛(wèi)生統(tǒng)計(jì)
3樓2015-10-19 09:05:11
4樓2015-10-19 12:02:43
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
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[考研] 293分求調(diào)劑，外語為俄語 +5 | 加一一九 2026-03-31 | 5/250 |
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[考研] 070300化學(xué)279求調(diào)劑 +6 | 哈哈哈^_^ 2026-03-31 | 8/400 |
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[考研] 一志愿華中師范化學(xué)332分求調(diào)劑 +3 | Lyy930824@ 2026-03-29 | 3/150 |
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[考研] 085600 材料與化工 329分求調(diào)劑 +18 | Mr. Z 2026-03-25 | 19/950 |
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[考研] 26考研-291分-廈門大學(xué)（085601）-柔性電子學(xué)院材料工程專業(yè)求調(diào)劑 +5 | min3 2026-03-24 | 6/300 |
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[考研] 332求調(diào)劑 +6 | @MZB382400 2026-03-28 | 6/300 |
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[考研] 0703 化學(xué) 求調(diào)劑，一志愿山東大學(xué) 342 分 +7 | Shern—- 2026-03-28 | 7/350 |
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[考研] 0856求調(diào)劑 +8 | 楒桉 2026-03-28 | 8/400 |
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[考研] 299求調(diào)劑 +10 | 15188958825 2026-03-25 | 10/500 |
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[考研] 085600，專業(yè)課化工原理，321分求調(diào)劑 +5 | 大饞小子 2026-03-28 | 5/250 |
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[考研] 11408軟件工程求調(diào)劑 +3 | Qiu學(xué)ing 2026-03-28 | 3/150 |
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[考研] 308求調(diào)劑 +7 | 墨墨漠 2026-03-27 | 7/350 |
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[考研] 272求調(diào)劑 +7 | 腳滑的守法公民 2026-03-27 | 7/350 |
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[考研] 08開頭275求調(diào)劑 +4 | 拉誰不重要 2026-03-26 | 4/200 |
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[論文投稿] Journal of Mechanical Science and Technology +3 | Russ_ss 2026-03-25 | 5/250 |
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[考研] 336材料求調(diào)劑 +7 | 陳瀅瑩 2026-03-26 | 9/450 |
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[考研] 總分322求生物學(xué)/生化與分子/生物信息學(xué)相關(guān)調(diào)劑 +5 | 星沉uu 2026-03-26 | 6/300 |
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[考研] 085601求調(diào)劑總分293英一數(shù)二 +4 | 鋼鐵大炮 2026-03-24 | 4/200 |
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[考研] 材料專碩 335 分求調(diào)劑 +4 | 拒絕冷暴力 2026-03-25 | 4/200 |
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[考研] 347求調(diào)劑 +4 | L when 2026-03-25 | 4/200 |
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