名称: | pLV3-H1-2O2-WDR55(human)-shRNA2-TetR-FLAG-Puro |
---|---|
别称: | |
基因长: | |
质粒宿主: | 哺乳细胞,慢病毒 |
质粒用途: | 干扰沉默,四环素诱导 |
片段类型: | shRNA |
片段物种: | 人 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | |
启动子: | H1-2O2 |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 37度 |
背景载体: | |
正向引物: | |
反向引物: | BGH:TAGAAGGCACAGTCGAGG |
诱导方式: |
1、本平台所有产品仅供科研使用,不能用于临床试验,包括人体口服、注射或者体外用药;
2、小包装质粒以模板价格出售,只保证基因部分测序正确,不保证质粒的表达等实验效果;
3、某些质粒即使是出品公司亦没有完整测过序,如关键部位的序列测序正确,亦认为是正确质粒;
4、小包装质粒产品的售后期为您收到产品之日起3个月,收到质粒后请及时转化操作,过期需重新购买;
5、质粒构建皆收取的是成本费用,所有质粒会分享给其他实验者,保密性质粒请提前告知并缴纳保密费用;
6、因科学研究为探索未知的活动,存在较大不确定性,故在任何情况下淼灵都不承担额外的连带责任;
7、发表[中文论文]请标注:产品名称(产品货号) 由武汉淼灵生物科技有限公司提供;
8、发表[英文论文]请标注:Product Name (Product Item Number) was obtained from MiaoLingBio,China.
操作说明
一、质粒干粉(负20度保存,保质期90天,常温运输)
1、收到干粉质粒后请先5000rpm离心1min,再加入20μl无菌水溶解质粒;
2、将相应的感受态于冰盒上解冻,取2μl质粒加至100μl感受态中,冰浴 30min;
3、42度热激60s,冰浴2min,加入900μl无抗LB培养基,180rpm震荡培养45min;
4、6000rpm离心3min,留100μl上清重悬,再涂至对应抗性的LB平板上;
5、倒置培养过夜后,如菌落过多则稀释质粒转化,如没有菌落则取10ul质粒转化;
6、挑单菌落至对应抗性的LB液体培养基中过夜震荡培养,根据需要提取质粒。
二、液体质粒(负20度保存,保质期90天,冰袋运输)
1、收到液体质粒后请甩动保存管,让液体质粒不要吸附在管壁上;
2、根据实验需求取适量质粒,其余保存于负20度冰箱。
三、甘油菌种(负80度保存,保质期90天,冰袋运输)
1、收到甘油菌种后请甩动保存管,让菌液不要吸附在管壁上;
2、用接种环在固体平板上四区划线,如有抗性就加对应抗性;
3、恒温培养箱倒置培养12~16h;
4、务必挑取单菌落接种于液体培养基中,如有抗性就加对应抗性;
5、摇床震荡培养过夜,根据实验需要培养菌种。
TTGGGGTTGC
GCCTTTTCCA
AGGCAGCCCT
GGGTTTGCGC
AGGGACGCGG
CTGCTCTGGG
CGTGGTTCCG
GGAAACGCAG
CGGCGCCGAC
CCTGGGTCTC
GCACATTCTT
CACGTCCGTT
CGCAGCGTCA
CCCGGATCTT
CGCCGCTACC
CTTGTGGGCC
CCCCGGCGAC
GCTTCCTGCT
CCGCCCCTAA
GTCGGGAAGG
TTCCTTGCGG
TTCGCGGCGT
GCCGGACGTG
ACAAACGGAA
GCCGCACGTC
TCACTAGTAC
CCTCGCAGAC
GGACAGCGCC
AGGGAGCAAT
GGCAGCGCGC
CGACCGCGAT
GGGCTGTGGC
CAATAGCGGC
TGCTCAGCAG
GGCGCGCCGA
GAGCAGCGGC
CGGGAAGGGG
CGGTGCGGGA
GGCGGGGTGT
GGGGCGGTAG
TGTGGGCCCT
GTTCCTGCCC
GCGCGGTGTT
CCGCATTCTG
CAAGCCTCCG
GAGCGCACGT
CGGCAGTCGG
CTCCCTCGTT
GACCGAATCA
CCGACCTCTC
TCCCCAGGGG
GATCTGTGAG
TTTGGGGACC
CTTGATTGTT
CTTTCTTTTT
CGCTATTGTA
AAATTCATGT
TATATGGAGG
GGGCAAAGTT
TTCAGGGTGT
TGTTTAGAAT
GGGAAGATGT
CCCTTGTATC
ACCATGGACC
CTCATGATAA
TTTTGTTTCT
TTCACTTTCT
ACTCTGTTGA
CAACCATTGT
CTCCTCTTAT
TTTCTTTTCA
TTTTCTGTAA
CTTTTTCGTT
AAACTTTAGC
TTGCATTTGT
AACGAATTTT
TAAATTCACT
TTTGTTTATT
TGTCAGATTG
TAAGTACTTT
CTCTAATCAC
TTTTTTTTCA
AGGCAATCAG
GGTATATTAT
ATTGTACTTC
AGCACAGTTT
TAGAGAACAA
TTGTTATAAT
TAAATGATAA
GGTAGAATAT
TTCTGCATAT
AAATTCTGGC
TGGCGTGGAA
ATATTCTTAT
TGGTAGAAAC
AACTACATCC
TGGTCATCAT
CCTGCCTTTC
TCTTTATGGT
TACAATGATA
TACACTGTTT
GAGATGAGGA
TAAAATACTC
TGAGTCCAAA
CCGGGCCCCT
CTGCTAACCA
TGTTCATGCC
TTCTTCTTTT
TCCTACAGCT
CCTGGGCAAC
GTGCTGGTTA
TTGTGCTGTC
TCATCATTTT
GGCAAAGAAT
TGTAATACGA
CTCACTATAG
GGCGAATTGA
TATGTCTAGA
TTAGATAAAA
GTAAAGTGAT
TAACAGCGCA
TTAGAGCTGC
TTAATGAGGT
CGGAATCGAA
GGTTTAACAA
CCCGTAAACT
CGCCCAGAAG
CTAGGTGTAG
AGCAGCCTAC
ATTGTATTGG
CATGTAAAAA
ATAAGCGGGC
TTTGCTCGAC
GCCTTAGCCA
TTGAGATGTT
AGATAGGCAC
CATACTCACT
TTTGCCCTTT
AGAAGGGGAA
AGCTGGCAAG
ATTTTTTACG
TAATAACGCT
AAAAGTTTTA
GATGTGCTTT
ACTAAGTCAT
CGCGATGGAG
CAAAAGTACA
TTTAGGTACA
CGGCCTACAG
AAAAACAGTA
TGAAACTCTC
GAAAATCAAT
TAGCCTTTTT
ATGCCAACAA
GGTTTTTCAC
TAGAGAATGC
ATTATATGCA
CTCAGCGCTG
TGGGGCATTT
TACTTTAGGT
TGCGTATTGG
AAGATCAAGA
GCATCAAGTC
GCTAAAGAAG
AAAGGGAAAC
ACCTACTACT
GATAGTATGC
CGCCATTATT
ACGACAAGCT
ATCGAATTAT
TTGATCACCA
AGGTGCAGAG
CCAGCCTTCT
TATTCGGCCT
TGAATTGATC
ATATGCGGAT
TAGAAAAACA
ACTTAAATGT
GAAAGTGGGT
CCGACTACAA
GGACGACGAT
GACAAGTAGC
CCGGGAGTTC
TAGGGATCTG
CCCCTCTCCC
TCCCCCCCCC
CTAACGTTAC
TGGCCGAAGC
CGCTTGGAAT
AAGGCCGGTG
TGCGTTTGTC
TATATGTTAT
TTTCCACCAT
ATTGCCGTCT
TTTGGCAATG
TGAGGGCCCG
GAAACCTGGC
CCTGTCTTCT
TGACGAGCAT
TCCTAGGGGT
CTTTCCCCTC
TCGCCAAAGG
AATGCAAGGT
CTGTTGAATG
TCGTGAAGGA
AGCAGTTCCT
CTGGAAGCTT
CTTGAAGACA
AACAACGTCT
GTAGCGACCC
TTTGCAGGCA
GCGGAACCCC
CCACCTGGCG
ACAGGTGCCT
CTGCGGCCAA
AAGCCACGTG
TATAAGATAC
ACCTGCAAAG
GCGGCACAAC
CCCAGTGCCA
CGTTGTGAGT
TGGATAGTTG
TGGAAAGAGT
CAAATGGCTC
TCCTCAAGCG
TATTCAACAA
GGGGCTGAAG
GATGCCCAGA
AGGTACCCCA
TTGTATGGGA
TCTGATCTGG
GGCCTCGGTG
CACATGCTTT
ACATGTGTTT
AGTCGAGGTT
AAAAAAACGT
CTAGGCCCCC
CGAACCACGG
GGACGTGGTT
TTCCTTTGAA
AAACACGATG
ATAAGGATCC
ACCGGAGCTT
ACCATGACCG
AGTACAAGCC
CACGGTGCGC
CTCGCCACCC
GCGACGACGT
CCCCAGGGCC
GTACGCACCC
TCGCCGCCGC
GTTCGCCGAC
TACCCCGCCA
CGCGCCACAC
CGTCGATCCG
GACCGCCACA
TCGAGCGGGT
CACCGAGCTG
CAAGAACTCT
TCCTCACGCG
CGTCGGGCTC
GACATCGGCA
AGGTGTGGGT
CGCGGACGAC
GGCGCCGCGG
TGGCGGTCTG
GACCACGCCG
GAGAGCGTCG
AAGCGGGGGC
GGTGTTCGCC
GAGATCGGCC
CGCGCATGGC
CGAGTTGAGC
GGTTCCCGGC
TGGCCGCGCA
GCAACAGATG
GAAGGCCTCC
TGGCGCCGCA
CCGGCCCAAG
GAGCCCGCGT
GGTTCCTGGC
CACCGTCGGC
GTCTCGCCCG
ACCACCAGGG
CAAGGGTCTG
GGCAGCGCCG
TCGTGCTCCC
CGGAGTGGAG
GCGGCCGAGC
GCGCCGGGGT
GCCCGCCTTC
CTGGAGACCT
CCGCGCCCCG
CAACCTCCCC
TTCTACGAGC
GGCTCGGCTT
CACCGTCACC
GCCGACGTCG
AGGTGCCCGA
AGGACCGCGC
ACCTGGTGCA
TGACCCGCAA
GCCCGGTGCC
TGACGCCCGC
CCCACGACCC
GCAGCGCCCG
ACCGAAAGGA
GCGCACGACC
CCATGCATCG
GTACCTTTAA
GACCAATGAC
TTACAAGGCA
GCTGTAGATC
TTAGCCACTT
TTTAAAAGAA
AAGGGGGGAC
TGGAAGGGCT
AATTCACTCC
CAACGAAGAC
AAGATCTGCT
TTTTGCTTGT
ACTGGGTCTC
TCTGGTTAGA
CCAGATCTGA
GCCTGGGAGC
TCTCTGGCTA
ACTAGGGAAC
CCACTGCTTA
AGCCTCAATA
AAGCTTGCCT
TGAGTGCTTC
AAGTAGTGTG
TGCCCGTCTG
TTGTGTGACT
CTGGTAACTA
GAGATCCCTC
AGACCCTTTT
AGTCAGTGTG
GAAAATCTCT
AGCAGTAGTA
GTTCATGTCA
TCTTATTATT
CAGTATTTAT
AACTTGCAAA
GAAATGAATA
TCAGAGAGTG
AGAGGAACTT
GTTTATTGCA
GCTTATAATG
GTTACAAATA
AAGCAATAGC
ATCACAAATT
TCACAAATAA
AGCATTTTTT
TCACTGCATT
CTAGTTGTGG
TTTGTCCAAA
CTCATCAATG
TATCTTATCA
TGTCTGGCTC
TAGCTATCCC
GCCCCTAACT
CCGCCCATCC
CGCCCCTAAC
TCCGCCCAGT
TCCGCCCATT
CTCCGCCCCA
TGGCTGACTA
ATTTTTTTTA
TTTATGCAGA
GGCCGAGGCC
GCCTCGGCCT
CTGAGCTATT
CCAGAAGTAG
TGAGGAGGCT
TTTTTGGAGG
CCTAGGGACG
TACCCAATTC
GCCCTATAGT
GAGTCGTATT
ACGCGCGCTC
ACTGGCCGTC
GTTTTACAAC
GTCGTGACTG
GGAAAACCCT
GGCGTTACCC
AACTTAATCG
CCTTGCAGCA
CATCCCCCTT
TCGCCAGCTG
GCGTAATAGC
GAAGAGGCCC
GCACCGATCG
CCCTTCCCAA
CAGTTGCGCA
GCCTGAATGG
CGAATGGGAC
GCGCCCTGTA
GCGGCGCATT
AAGCGCGGCG
GGTGTGGTGG
TTACGCGCAG
CGTGACCGCT
ACACTTGCCA
GCGCCCTAGC
GCCCGCTCCT
TTCGCTTTCT
TCCCTTCCTT
TCTCGCCACG
TTCGCCGGCT
TTCCCCGTCA
AGCTCTAAAT
CGGGGGCTCC
CTTTAGGGTT
CCGATTTAGT
GCTTTACGGC
ACCTCGACCC
CAAAAAACTT
GATTAGGGTG
ATGGTTCACG
TAGTGGGCCA
TCGCCCTGAT
AGACGGTTTT
TCGCCCTTTG
ACGTTGGAGT
CCACGTTCTT
TAATAGTGGA
CTCTTGTTCC
AAACTGGAAC
AACACTCAAC
CCTATCTCGG
TCTATTCTTT
TGATTTATAA
GGGATTTTGC
CGATTTCGGC
CTATTGGTTA
AAAAATGAGC
TGATTTAACA
AAAATTTAAC
GCGAATTTTA
ACAAAATATT
AACGCTTACA
ATTTAGGTGG
CACTTTTCGG
GGAAATGTGC
GCGGAACCCC
TATTTGTTTA
TTTTTCTAAA
TACATTCAAA
TATGTATCCG
CTCATGAGAC
AATAACCCTG
ATAAATGCTT
CAATAATATT
GAAAAAGGAA
GAGTATGAGT
ATTCAACATT
TCCGTGTCGC
CCTTATTCCC
TTTTTTGCGG
CATTTTGCCT
TCCTGTTTTT
GCTCACCCAG
AAACGCTGGT
GAAAGTAAAA
GATGCTGAAG
ATCAGTTGGG
TGCACGAGTG
GGTTACATCG
AACTGGATCT
CAACAGCGGT
AAGATCCTTG
AGAGTTTTCG
CCCCGAAGAA
CGTTTTCCAA
TGATGAGCAC
TTTTAAAGTT
CTGCTATGTG
GCGCGGTATT
ATCCCGTATT
GACGCCGGGC
AAGAGCAACT
CGGTCGCCGC
ATACACTATT
CTCAGAATGA
CTTGGTTGAG
TACTCACCAG
TCACAGAAAA
GCATCTTACG
GATGGCATGA
CAGTAAGAGA
ATTATGCAGT
GCTGCCATAA
CCATGAGTGA
TAACACTGCG
GCCAACTTAC
TTCTGACAAC
GATCGGAGGA
CCGAAGGAGC
TAACCGCTTT
TTTGCACAAC
ATGGGGGATC
ATGTAACTCG
CCTTGATCGT
TGGGAACCGG
AGCTGAATGA
AGCCATACCA
AACGACGAGC
GTGACACCAC
GATGCCTGTA
GCAATGGCAA
CAACGTTGCG
CAAACTATTA
ACTGGCGAAC
TACTTACTCT
AGCTTCCCGG
CAACAATTAA
TAGACTGGAT
GGAGGCGGAT
AAAGTTGCAG
GACCACTTCT
GCGCTCGGCC
CTTCCGGCTG
GCTGGTTTAT
TGCTGATAAA
TCTGGAGCCG
GTGAGCGTGG
GTCTCGCGGT
ATCATTGCAG
CACTGGGGCC
AGATGGTAAG
CCCTCCCGTA
TCGTAGTTAT
CTACACGACG
GGGAGTCAGG
CAACTATGGA
TGAACGAAAT
AGACAGATCG
CTGAGATAGG
TGCCTCACTG
ATTAAGCATT
GGTAACTGTC
AGACCAAGTT
TACTCATATA
TACTTTAGAT
TGATTTAAAA
CTTCATTTTT
AATTTAAAAG
GATCTAGGTG
AAGATCCTTT
TTGATAATCT
CATGACCAAA
ATCCCTTAAC
GTGAGTTTTC
GTTCCACTGA
GCGTCAGACC
CCGTAGAAAA
GATCAAAGGA
TCTTCTTGAG
ATCCTTTTTT
TCTGCGCGTA
ATCTGCTGCT
TGCAAACAAA
AAAACCACCG
CTACCAGCGG
TGGTTTGTTT
GCCGGATCAA
GAGCTACCAA
CTCTTTTTCC
GAAGGTAACT
GGCTTCAGCA
GAGCGCAGAT
ACCAAATACT
GTTCTTCTAG
TGTAGCCGTA
GTTAGGCCAC
CACTTCAAGA
ACTCTGTAGC
ACCGCCTACA
TACCTCGCTC
TGCTAATCCT
GTTACCAGTG
GCTGCTGCCA
GTGGCGATAA
GTCGTGTCTT
ACCGGGTTGG
ACTCAAGACG
ATAGTTACCG
GATAAGGCGC
AGCGGTCGGG
CTGAACGGGG
GGTTCGTGCA
CACAGCCCAG
CTTGGAGCGA
ACGACCTACA
CCGAACTGAG
ATACCTACAG
CGTGAGCTAT
GAGAAAGCGC
CACGCTTCCC
GAAGGGAGAA
AGGCGGACAG
GTATCCGGTA
AGCGGCAGGG
TCGGAACAGG
AGAGCGCACG
AGGGAGCTTC
CAGGGGGAAA
CGCCTGGTAT
CTTTATAGTC
CTGTCGGGTT
TCGCCACCTC
TGACTTGAGC
GTCGATTTTT
GTGATGCTCG
TCAGGGGGGC
GGAGCCTATG
GAAAAACGCC
AGCAACGCGG
CCTTTTTACG
GTTCCTGGCC
TTTTGCTGGC
CTTTTGCTCA
CATGTTCTTT
CCTGCGTTAT
CCCCTGATTC
TGTGGATAAC
CGTATTACCG
CCTTTGAGTG
AGCTGATACC
GCTCGCCGCA
GCCGAACGAC
CGAGCGCAGC
GAGTCAGTGA
GCGAGGAAGC
GGAAGAGCGC
CCAATACGCA
AACCGCCTCT
CCCCGCGCGT
TGGCCGATTC
ATTAATGCAG
CTGGCACGAC
AGGTTTCCCG
ACTGGAAAGC
GGGCAGTGAG
CGCAACGCAA
TTAATGTGAG
TTAGCTCACT
CATTAGGCAC
CCCAGGCTTT
ACACTTTATG
CTTCCGGCTC
GTATGTTGTG
TGGAATTGTG
AGCGGATAAC
AATTTCACAC
AGGAAACAGC
TATGACCATG
ATTACGCCAA
GCGCGCAATT
AACCCTCACT
AAAGGGAACA
AAAGCTGGAG
CTGCAAGCTT
AATGTAGTCT
TATGCAATAC
TCTTGTAGTC
TTGCAACATG
GTAACGATGA
GTTAGCAACA
TGCCTTACAA
GGAGAGAAAA
AGCACCGTGC
ATGCCGATTG
GTGGAAGTAA
GGTGGTACGA
TCGTGCCTTA
TTAGGAAGGC
AACAGACGGG
TCTGACATGG
ATTGGACGAA
CCACTGAATT
GCCGCATTGC
AGAGATATTG
TATTTAAGTG
CCTAGCTCGA
TACATAAACG
GGTCTCTCTG
GTTAGACCAG
ATCTGAGCCT
GGGAGCTCTC
TGGCTAACTA
GGGAACCCAC
TGCTTAAGCC
TCAATAAAGC
TTGCCTTGAG
TGCTTCAAGT
AGTGTGTGCC
CGTCTGTTGT
GTGACTCTGG
TAACTAGAGA
TCCCTCAGAC
CCTTTTAGTC
AGTGTGGAAA
ATCTCTAGCA
GTGGCGCCCG
AACAGGGACT
TGAAAGCGAA
AGGGAAACCA
GAGGAGCTCT
CTCGACGCAG
GACTCGGCTT
GCTGAAGCGC
GCACGGCAAG
AGGCGAGGGG
CGGCGACTGG
TGAGTACGCC
AAAAATTTTG
ACTAGCGGAG
GCTAGAAGGA
GAGAGATGGG
TGCGAGAGCG
TCAGTATTAA
GCGGGGGAGA
ATTAGATCGC
GATGGGAAAA
AATTCGGTTA
AGGCCAGGGG
GAAAGAAAAA
ATATAAATTA
AAACATATAG
TATGGGCAAG
CAGGGAGCTA
GAACGATTCG
CAGTTAATCC
TGGCCTGTTA
GAAACATCAG
AAGGCTGTAG
ACAAATACTG
GGACAGCTAC
AACCATCCCT
TCAGACAGGA
TCAGAAGAAC
TTAGATCATT
ATATAATACA
GTAGCAACCC
TCTATTGTGT
GCATCAAAGG
ATAGAGATAA
AAGACACCAA
GGAAGCTTTA
GACAAGATAG
AGGAAGAGCA
AAACAAAAGT
AAGACCACCG
CACAGCAAGC
GGCCGCTGAT
CTTCAGACCT
GGAGGAGGAG
ATATGAGGGA
CAATTGGAGA
AGTGAATTAT
ATAAATATAA
AGTAGTAAAA
ATTGAACCAT
TAGGAGTAGC
ACCCACCAAG
GCAAAGAGAA
GAGTGGTGCA
GAGAGAAAAA
AGAGCAGTGG
GAATAGGAGC
TTTGTTCCTT
GGGTTCTTGG
GAGCAGCAGG
AAGCACTATG
GGCGCAGCGT
CAATGACGCT
GACGGTACAG
GCCAGACAAT
TATTGTCTGG
TATAGTGCAG
CAGCAGAACA
ATTTGCTGAG
GGCTATTGAG
GCGCAACAGC
ATCTGTTGCA
ACTCACAGTC
TGGGGCATCA
AGCAGCTCCA
GGCAAGAATC
CTGGCTGTGG
AAAGATACCT
AAAGGATCAA
CAGCTCCTGG
GGATTTGGGG
TTGCTCTGGA
AAACTCATTT
GCACCACTGC
TGTGCCTTGG
AATGCTAGTT
GGAGTAATAA
ATCTCTGGAA
CAGATTTGGA
ATCACACGAC
CTGGATGGAG
TGGGACAGAG
AAATTAACAA
TTACACAAGC
TTAATACACT
CCTTAATTGA
AGAATCGCAA
AACCAGCAAG
AAAAGAATGA
ACAAGAATTA
TTGGAATTAG
ATAAATGGGC
AAGTTTGTGG
AATTGGTTTA
ACATAACAAA
TTGGCTGTGG
TATATAAAAT
TATTCATAAT
GATAGTAGGA
GGCTTGGTAG
GTTTAAGAAT
AGTTTTTGCT
GTACTTTCTA
TAGTGAATAG
AGTTAGGCAG
GGATATTCAC
CATTATCGTT
TCAGACCCAC
CTCCCAACCC
CGAGGGGACC
CGACAGGCCC
GAAGGAATAG
AAGAAGAAGG
TGGAGAGAGA
GACAGAGACA
GATCCATTCG
ATTAGTGAAC
GGATCTCGAC
GGTATCGATC
ACGAGACTAG
CCTCGAGCGG
CCGCAATATT
TGCATGTCGC
TATGTGTTCT
GGGAAATCAC
CATAAACGTG
AAATCCCTAT
CAGTGATAGA
GACTTATAAG
TTCCCTATCA
GTGATAGAGA
CACCGGGTAC
CATCTACCTC
TTCAATTCTC
GAGAATTGAA
GAGGTAGATG
GTACTTTTTT
GAATTCTCGA
CCTCGAGACA
AATGGCAGTA
TTCATCCACA
ATTTTAAAAG
AAAAGGGGGG
ATTGGGGGGT
ACAGTGCAGG
GGAAAGAATA
GTAGACATAA
TAGCAACAGA
CATACAAACT
AAAGAATTAC
AAAAACAAAT
TACAAAAATT
CAAAATTTTC
GGGTTTATTA
CAGGGACAGC
AGAGATCCAC
TTTGGCCGCG
GCTCGAGGGG
G