名称: | pAC152-dual-dCas9VP64-sgExpression |
---|---|
别称: | Plasmid#48238 |
基因长: | |
质粒宿主: | 哺乳细胞 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR |
片段物种: | |
原核抗性: | Amp |
筛选标记: | |
荧光标记: | |
启动子: | |
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: | |
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。
GAGGGCCTAT
TTCCCATGAT
TCCTTCATAT
TTGCATATAC
GATACAAGGC
TGTTAGAGAG
ATAATTGGAA
TTAATTTGAC
TGTAAACACA
AAGATATTAG
TACAAAATAC
GTGACGTAGA
AAGTAATAAT
TTCTTGGGTA
GTTTGCAGTT
TTAAAATTAT
GTTTTAAAAT
GGACTATCAT
ATGCTTACCG
TAACTTGAAA
GTATTTCGAT
TTCTTGGCTT
TATATATCTT
GTGGAAAGGA
CGAAACACCG
GGTCTTCGAG
AAGACCTGTT
TTAGAGCTAG
AAATAGCAAG
TTAAAATAAG
GCTAGTCCGT
TATCAACTTG
AAAAAGTGGC
ACCGAGTCGG
TGCTTTTTTG
TTTTAGAGCT
AGAAATAGCA
AGTTAAAATA
AGGCTAGTCC
GTTTTTAGCG
CGTGCGCCAA
TTCTGCAGAC
AAATGGCTCT
AGAGGTACCC
GTTACATAAC
TTACGGTAAA
TGGCCCGCCT
GGCTGACCGC
CCAACGACCC
CCGCCCATTG
ACGTCAATAG
TAACGCCAAT
AGGGACTTTC
CATTGACGTC
AATGGGTGGA
GTATTTACGG
TAAACTGCCC
ACTTGGCAGT
ACATCAAGTG
TATCATATGC
CAAGTACGCC
CCCTATTGAC
GTCAATGACG
GTAAATGGCC
CGCCTGGCAT
TGTGCCCAGT
ACATGACCTT
ATGGGACTTT
CCTACTTGGC
AGTACATCTA
CGTATTAGTC
ATCGCTATTA
CCATGGTCGA
GGTGAGCCCC
ACGTTCTGCT
TCACTCTCCC
CATCTCCCCC
CCCTCCCCAC
CCCCAATTTT
GTATTTATTT
ATTTTTTAAT
TATTTTGTGC
AGCGATGGGG
GCGGGGGGGG
GGGGGGGGCG
CGCGCCAGGC
GGGGCGGGGC
GGGGCGAGGG
GCGGGGCGGG
GCGAGGCGGA
GAGGTGCGGC
GGCAGCCAAT
CAGAGCGGCG
CGCTCCGAAA
GTTTCCTTTT
ATGGCGAGGC
GGCGGCGGCG
GCGGCCCTAT
AAAAAGCGAA
GCGCGCGGCG
GGCGGGAGTC
GCTGCGACGC
TGCCTTCGCC
CCGTGCCCCG
CTCCGCCGCC
GCCTCGCGCC
GCCCGCCCCG
GCTCTGACTG
ACCGCGTTAC
TCCCACAGGT
GAGCGGGCGG
GACGGCCCTT
CTCCTCCGGG
CTGTAATTAG
CTGAGCAAGA
GGTAAGGGTT
TAAGGGATGG
TTGGTTGGTG
GGGTATTAAT
GTTTAATTAC
CTGGAGCACC
TGCCTGAAAT
CACTTTTTTT
CAGGTTGGAC
CGGTGCCACC
ATGTACCCAT
ACGATGTTCC
AGATTACGCT
TCGCCGAAGA
AAAAGCGCAA
GGTCGAAGCG
TCCGACAAGA
AGTACAGCAT
CGGCCTGGCC
ATCGGCACCA
ACTCTGTGGG
CTGGGCCGTG
ATCACCGACG
AGTACAAGGT
GCCCAGCAAG
AAATTCAAGG
TGCTGGGCAA
CACCGACCGG
CACAGCATCA
AGAAGAACCT
GATCGGAGCC
CTGCTGTTCG
ACAGCGGCGA
AACAGCCGAG
GCCACCCGGC
TGAAGAGAAC
CGCCAGAAGA
AGATACACCA
GACGGAAGAA
CCGGATCTGC
TATCTGCAAG
AGATCTTCAG
CAACGAGATG
GCCAAGGTGG
ACGACAGCTT
CTTCCACAGA
CTGGAAGAGT
CCTTCCTGGT
GGAAGAGGAT
AAGAAGCACG
AGCGGCACCC
CATCTTCGGC
AACATCGTGG
ACGAGGTGGC
CTACCACGAG
AAGTACCCCA
CCATCTACCA
CCTGAGAAAG
AAACTGGTGG
ACAGCACCGA
CAAGGCCGAC
CTGCGGCTGA
TCTATCTGGC
CCTGGCCCAC
ATGATCAAGT
TCCGGGGCCA
CTTCCTGATC
GAGGGCGACC
TGAACCCCGA
CAACAGCGAC
GTGGACAAGC
TGTTCATCCA
GCTGGTGCAG
ACCTACAACC
AGCTGTTCGA
GGAAAACCCC
ATCAACGCCA
GCGGCGTGGA
CGCCAAGGCC
ATCCTGTCTG
CCAGACTGAG
CAAGAGCAGA
CGGCTGGAAA
ATCTGATCGC
CCAGCTGCCC
GGCGAGAAGA
AGAATGGCCT
GTTCGGCAAC
CTGATTGCCC
TGAGCCTGGG
CCTGACCCCC
AACTTCAAGA
GCAACTTCGA
CCTGGCCGAG
GATGCCAAAC
TGCAGCTGAG
CAAGGACACC
TACGACGACG
ACCTGGACAA
CCTGCTGGCC
CAGATCGGCG
ACCAGTACGC
CGACCTGTTT
CTGGCCGCCA
AGAACCTGTC
CGACGCCATC
CTGCTGAGCG
ACATCCTGAG
AGTGAACACC
GAGATCACCA
AGGCCCCCCT
GAGCGCCTCT
ATGATCAAGA
GATACGACGA
GCACCACCAG
GACCTGACCC
TGCTGAAAGC
TCTCGTGCGG
CAGCAGCTGC
CTGAGAAGTA
CAAAGAGATT
TTCTTCGACC
AGAGCAAGAA
CGGCTACGCC
GGCTACATTG
ACGGCGGAGC
CAGCCAGGAA
GAGTTCTACA
AGTTCATCAA
GCCCATCCTG
GAAAAGATGG
ACGGCACCGA
GGAACTGCTC
GTGAAGCTGA
ACAGAGAGGA
CCTGCTGCGG
AAGCAGCGGA
CCTTCGACAA
CGGCAGCATC
CCCCACCAGA
TCCACCTGGG
AGAGCTGCAC
GCCATTCTGC
GGCGGCAGGA
AGATTTTTAC
CCATTCCTGA
AGGACAACCG
GGAAAAGATC
GAGAAGATCC
TGACCTTCCG
CATCCCCTAC
TACGTGGGCC
CTCTGGCCAG
GGGAAACAGC
AGATTCGCCT
GGATGACCAG
AAAGAGCGAG
GAAACCATCA
CCCCCTGGAA
CTTCGAGGAA
GTGGTGGACA
AGGGCGCTTC
CGCCCAGAGC
TTCATCGAGC
GGATGACCAA
CTTCGATAAG
AACCTGCCCA
ACGAGAAGGT
GCTGCCCAAG
CACAGCCTGC
TGTACGAGTA
CTTCACCGTG
TATAACGAGC
TGACCAAAGT
GAAATACGTG
ACCGAGGGAA
TGAGAAAGCC
CGCCTTCCTG
AGCGGCGAGC
AGAAAAAGGC
CATCGTGGAC
CTGCTGTTCA
AGACCAACCG
GAAAGTGACC
GTGAAGCAGC
TGAAAGAGGA
CTACTTCAAG
AAAATCGAGT
GCTTCGACTC
CGTGGAAATC
TCCGGCGTGG
AAGATCGGTT
CAACGCCTCC
CTGGGCACAT
ACCACGATCT
GCTGAAAATT
ATCAAGGACA
AGGACTTCCT
GGACAATGAG
GAAAACGAGG
ACATTCTGGA
AGATATCGTG
CTGACCCTGA
CACTGTTTGA
GGACAGAGAG
ATGATCGAGG
AACGGCTGAA
AACCTATGCC
CACCTGTTCG
ACGACAAAGT
GATGAAGCAG
CTGAAGCGGC
GGAGATACAC
CGGCTGGGGC
AGGCTGAGCC
GGAAGCTGAT
CAACGGCATC
CGGGACAAGC
AGTCCGGCAA
GACAATCCTG
GATTTCCTGA
AGTCCGACGG
CTTCGCCAAC
AGAAACTTCA
TGCAGCTGAT
CCACGACGAC
AGCCTGACCT
TTAAAGAGGA
CATCCAGAAA
GCCCAGGTGT
CCGGCCAGGG
CGATAGCCTG
CACGAGCACA
TTGCCAATCT
GGCCGGCAGC
CCCGCCATTA
AGAAGGGCAT
CCTGCAGACA
GTGAAGGTGG
TGGACGAGCT
CGTGAAAGTG
ATGGGCCGGC
ACAAGCCCGA
GAACATCGTG
ATCGAAATGG
CCAGAGAGAA
CCAGACCACC
CAGAAGGGAC
AGAAGAACAG
CCGCGAGAGA
ATGAAGCGGA
TCGAAGAGGG
CATCAAAGAG
CTGGGCAGCC
AGATCCTGAA
AGAACACCCC
GTGGAAAACA
CCCAGCTGCA
GAACGAGAAG
CTGTACCTGT
ACTACCTGCA
GAATGGGCGG
GATATGTACG
TGGACCAGGA
ACTGGACATC
AACCGGCTGT
CCGACTACGA
TGTGGACGCC
ATCGTGCCTC
AGAGCTTTCT
GAAGGACGAC
TCCATCGACA
ACAAGGTGCT
GACCAGAAGC
GACAAGAACC
GGGGCAAGAG
CGACAACGTG
CCCTCCGAAG
AGGTCGTGAA
GAAGATGAAG
AACTACTGGC
GGCAGCTGCT
GAACGCCAAG
CTGATTACCC
AGAGAAAGTT
CGACAATCTG
ACCAAGGCCG
AGAGAGGCGG
CCTGAGCGAA
CTGGATAAGG
CCGGCTTCAT
CAAGAGACAG
CTGGTGGAAA
CCCGGCAGAT
CACAAAGCAC
GTGGCACAGA
TCCTGGACTC
CCGGATGAAC
ACTAAGTACG
ACGAGAATGA
CAAGCTGATC
CGGGAAGTGA
AAGTGATCAC
CCTGAAGTCC
AAGCTGGTGT
CCGATTTCCG
GAAGGATTTC
CAGTTTTACA
AAGTGCGCGA
GATCAACAAC
TACCACCACG
CCCACGACGC
CTACCTGAAC
GCCGTCGTGG
GAACCGCCCT
GATCAAAAAG
TACCCTAAGC
TGGAAAGCGA
GTTCGTGTAC
GGCGACTACA
AGGTGTACGA
CGTGCGGAAG
ATGATCGCCA
AGAGCGAGCA
GGAAATCGGC
AAGGCTACCG
CCAAGTACTT
CTTCTACAGC
AACATCATGA
ACTTTTTCAA
GACCGAGATT
ACCCTGGCCA
ACGGCGAGAT
CCGGAAGCGG
CCTCTGATCG
AGACAAACGG
CGAAACCGGG
GAGATCGTGT
GGGATAAGGG
CCGGGATTTT
GCCACCGTGC
GGAAAGTGCT
GAGCATGCCC
CAAGTGAATA
TCGTGAAAAA
GACCGAGGTG
CAGACAGGCG
GCTTCAGCAA
AGAGTCTATC
CTGCCCAAGA
GGAACAGCGA
TAAGCTGATC
GCCAGAAAGA
AGGACTGGGA
CCCTAAGAAG
TACGGCGGCT
TCGACAGCCC
CACCGTGGCC
TATTCTGTGC
TGGTGGTGGC
CAAAGTGGAA
AAGGGCAAGT
CCAAGAAACT
GAAGAGTGTG
AAAGAGCTGC
TGGGGATCAC
CATCATGGAA
AGAAGCAGCT
TCGAGAAGAA
TCCCATCGAC
TTTCTGGAAG
CCAAGGGCTA
CAAAGAAGTG
AAAAAGGACC
TGATCATCAA
GCTGCCTAAG
TACTCCCTGT
TCGAGCTGGA
AAACGGCCGG
AAGAGAATGC
TGGCCTCTGC
CGGCGAACTG
CAGAAGGGAA
ACGAACTGGC
CCTGCCCTCC
AAATATGTGA
ACTTCCTGTA
CCTGGCCAGC
CACTATGAGA
AGCTGAAGGG
CTCCCCCGAG
GATAATGAGC
AGAAACAGCT
GTTTGTGGAA
CAGCACAAGC
ACTACCTGGA
CGAGATCATC
GAGCAGATCA
GCGAGTTCTC
CAAGAGAGTG
ATCCTGGCCG
ACGCTAATCT
GGACAAAGTG
CTGTCCGCCT
ACAACAAGCA
CCGGGATAAG
CCCATCAGAG
AGCAGGCCGA
GAATATCATC
CACCTGTTTA
CCCTGACCAA
TCTGGGAGCC
CCTGCCGCCT
TCAAGTACTT
TGACACCACC
ATCGACCGGA
AGAGGTACAC
CAGCACCAAA
GAGGTGCTGG
ACGCCACCCT
GATCCACCAG
AGCATCACCG
GCCTGTACGA
GACACGGATC
GACCTGTCTC
AGCTGGGAGG
CGACAGCCCC
AAGAAGAAGA
GAAAGGTGGA
GGCCAGCGGG
CCGGCCGGAT
CCGGGCGCGC
CGACGCGCTG
GACGATTTCG
ATCTCGACAT
GCTGGGTTCT
GATGCCCTCG
ATGACTTTGA
CCTGGATATG
TTGGGAAGCG
ACGCATTGGA
TGACTTTGAT
CTGGACATGC
TCGGCTCCGA
TGCTCTGGAC
GATTTCGATC
TCGATATGTT
ATATATCGAT
TGATTAATTA
AGAATTCCTA
GAGCTCGCTG
ATCAGCCTCG
ACTGTGCCTT
CTAGTTGCCA
GCCATCTGTT
GTTTGCCCCT
CCCCCGTGCC
TTCCTTGACC
CTGGAAGGTG
CCACTCCCAC
TGTCCTTTCC
TAATAAAATG
AGGAAATTGC
ATCGCATTGT
CTGAGTAGGT
GTCATTCTAT
TCTGGGGGGT
GGGGTGGGGC
AGGACAGCAA
GGGGGAGGAT
TGGGAAGAGA
ATAGCAGGCA
TGCTGGGGAG
CGGCCGCAGG
AACCCCTAGT
GATGGAGTTG
GCCACTCCCT
CTCTGCGCGC
TCGCTCGCTC
ACTGAGGCCG
GGCGACCAAA
GGTCGCCCGA
CGCCCGGGCT
TTGCCCGGGC
GGCCTCAGTG
AGCGAGCGAG
CGCGCAGCTG
CCTGCAGGGG
CGCCTGATGC
GGTATTTTCT
CCTTACGCAT
CTGTGCGGTA
TTTCACACCG
CATACGTCAA
AGCAACCATA
GTACGCGCCC
TGTAGCGGCG
CATTAAGCGC
GGCGGGTGTG
GTGGTTACGC
GCAGCGTGAC
CGCTACACTT
GCCAGCGCCC
TAGCGCCCGC
TCCTTTCGCT
TTCTTCCCTT
CCTTTCTCGC
CACGTTCGCC
GGCTTTCCCC
GTCAAGCTCT
AAATCGGGGG
CTCCCTTTAG
GGTTCCGATT
TAGTGCTTTA
CGGCACCTCG
ACCCCAAAAA
ACTTGATTTG
GGTGATGGTT
CACGTAGTGG
GCCATCGCCC
TGATAGACGG
TTTTTCGCCC
TTTGACGTTG
GAGTCCACGT
TCTTTAATAG
TGGACTCTTG
TTCCAAACTG
GAACAACACT
CAACCCTATC
TCGGGCTATT
CTTTTGATTT
ATAAGGGATT
TTGCCGATTT
CGGCCTATTG
GTTAAAAAAT
GAGCTGATTT
AACAAAAATT
TAACGCGAAT
TTTAACAAAA
TATTAACGTT
TACAATTTTA
TGGTGCACTC
TCAGTACAAT
CTGCTCTGAT
GCCGCATAGT
TAAGCCAGCC
CCGACACCCG
CCAACACCCG
CTGACGCGCC
CTGACGGGCT
TGTCTGCTCC
CGGCATCCGC
TTACAGACAA
GCTGTGACCG
TCTCCGGGAG
CTGCATGTGT
CAGAGGTTTT
CACCGTCATC
ACCGAAACGC
GCGAGACGAA
AGGGCCTCGT
GATACGCCTA
TTTTTATAGG
TTAATGTCAT
GATAATAATG
GTTTCTTAGA
CGTCAGGTGG
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
AAGCCGCGGT
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
GTCCTTCTAG
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
CATGT