名称: | pCMV-T7-SpCas9-NG-P2A-EGFP (RTW3677)② |
---|---|
别称: | Plasmid#139994② |
基因长: | |
质粒宿主: | 哺乳细胞 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR,Cas9 |
片段物种: | |
原核抗性: | Amp |
筛选标记: | |
荧光标记: | 绿色 |
启动子: | CMV |
复制子: | pUC |
拷贝数: | |
感受态: | DH5a |
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。
GAGGTCGCTG
AGTAGTGCGC
GAGCAAAATT
TAAGCTACAA
CAAGGCAAGG
CTTGACCGAC
AATTGCATGA
AGAATCTGCT
TAGGGTTAGG
CGTTTTGCGC
TGCTTCGCGA
TGTACGGGCC
AGATATACGC
GTTGACATTG
ATTATTGACT
AGTTATTAAT
AGTAATCAAT
TACGGGGTCA
TTAGTTCATA
GCCCATATAT
GGAGTTCCGC
GTTACATAAC
TTACGGTAAA
TGGCCCGCCT
GGCTGACCGC
CCAACGACCC
CCGCCCATTG
ACGTCAATAA
TGACGTATGT
TCCCATAGTA
ACGCCAATAG
GGACTTTCCA
TTGACGTCAA
TGGGTGGAGT
ATTTACGGTA
AACTGCCCAC
TTGGCAGTAC
ATCAAGTGTA
TCATATGCCA
AGTACGCCCC
CTATTGACGT
CAATGACGGT
AAATGGCCCG
CCTGGCATTA
TGCCCAGTAC
ATGACCTTAT
GGGACTTTCC
TACTTGGCAG
TACATCTACG
TATTAGTCAT
CGCTATTACC
ATGGTGATGC
GGTTTTGGCA
GTACATCAAT
GGGCGTGGAT
AGCGGTTTGA
CTCACGGGGA
TTTCCAAGTC
TCCACCCCAT
TGACGTCAAT
GGGAGTTTGT
TTTGGCACCA
AAATCAACGG
GACTTTCCAA
AATGTCGTAA
CAACTCCGCC
CCATTGACGC
AAATGGGCGG
TAGGCGTGTA
CGGTGGGAGG
TCTATATAAG
CAGAGCTGGT
TTAGTGAACC
GTCAGATCCG
CTAGAGATCC
GCGGCCGCTA
ATACGACTCA
CTATAGGGAG
AGCCGCCACC
ATGGACAAGA
AGTACAGCAT
CGGCCTGGAC
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
GTTCGGAAAC
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
TGTGGACCAT
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
AGACCCAAGA
GGAACAGCGA
TAAGCTGATC
GCCAGAAAGA
AGGACTGGGA
CCCTAAGAAG
TACGGCGGCT
TCGTGAGCCC
CACCGTGGCC
TATTCTGTGC
TGGTGGTGGC
CAAAGTGGAA
AAGGGCAAGT
CCAAGAAACT
GAAGAGTGTG
AAAGAGCTGC
TGGGGATCAC
CATCATGGAA
AGAAGCAGCT
TCGAGAAGAA
TCCCATCGAC
TTTCTGGAAG
CCAAGGGCTA
CAAAGAAGTG
AAAAAGGACC
TGATCATCAA
GCTGCCTAAG
TACTCCCTGT
TCGAGCTGGA
AAACGGCCGG
AAGAGAATGC
TGGCCTCTGC
CAGATTCCTG
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
CCTAGAGCCT
TCAAGTACTT
TGACACCACC
ATCGACCGGA
AGGTGTACAG
AAGCACCAAA
GAGGTGCTGG
ACGCCACCCT
GATCCACCAG
AGCATCACCG
GCCTGTACGA
GACACGGATC
GACCTGTCTC
AGCTGGGAGG
CGACGGATCC
GGCGGAGGCG
GAAGCGGGAA
AAGAACCGCC
GACGGCAGCG
AATTCGAGCC
CAAGAAGAAG
AGGAAAGTCT
CGAGCGGAGG
CGACTACAAA
GACCATGACG
GTGATTATAA
AGATCATGAC
ATCGATTACA
AGGATGACGA
TGACAAGGGA
GGCGGTGGAA
GCGGCGCAAC
AAACTTCTCT
CTGCTGAAAC
AAGCCGGAGA
TGTCGAAGAG
AATCCTGGAC
CGATGGTGAG
CAAGGGCGAG
GAGCTGTTCA
CCGGGGTGGT
GCCCATCCTG
GTCGAGCTGG
ACGGCGACGT
AAACGGCCAC
AAGTTCAGCG
TGTCCGGCGA
GGGCGAGGGC
GATGCCACCT
ACGGCAAGCT
GACCCTGAAG
TTCATCTGCA
CCACCGGCAA
GCTGCCCGTG
CCCTGGCCCA
CCCTCGTGAC
CACCCTGACC
TACGGCGTGC
AGTGCTTCAG
CCGCTACCCC
GACCACATGA
AGCAGCACGA
CTTCTTCAAG
TCCGCCATGC
CCGAAGGCTA
CGTCCAGGAG
CGCACCATCT
TCTTCAAGGA
CGACGGCAAC
TACAAGACCC
GCGCCGAGGT
GAAGTTCGAG
GGCGACACCC
TGGTGAACCG
CATCGAGCTG
AAGGGCATCG
ACTTCAAGGA
GGACGGCAAC
ATCCTGGGGC
ACAAGCTGGA
GTACAACTAC
AACAGCCACA
ACGTCTATAT
CATGGCCGAC
AAGCAGAAGA
ACGGCATCAA
GGTGAACTTC
AAGATCCGCC
ACAACATCGA
GGACGGCAGC
GTGCAGCTCG
CCGACCACTA
CCAGCAGAAC
ACCCCCATCG
GCGACGGCCC
CGTGCTGCTG
CCCGACAACC
ACTACCTGAG
CACCCAGTCC
GCCCTGAGCA
AAGACCCCAA
CGAGAAGCGC
GATCACATGG
TCCTGCTGGA
GTTCGTGACC
GCCGCCGGGA
TCACTCTCGG
CATGGACGAG
CTGTACAAGT
GACCGGTCAT
CATCACCATC
ACCATTGAGT
TTAAACCCGC
TGATCAGCCT
CGACTGTGCC
TTCTAGTTGC
CAGCCATCTG
TTGTTTGCCC
CTCCCCCGTG
CCTTCCTTGA
CCCTGGAAGG
TGCCACTCCC
ACTGTCCTTT
CCTAATAAAA
TGAGGAAATT
GCATCGCATT
GTCTGAGTAG
GTGTCATTCT
ATTCTGGGGG
GTGGGGTGGG
GCAGGACAGC
AAGGGGGAGG
ATTGGGAAGA
CAATAGCAGG
CATGCTGGGG
ATGCGGTGGG
CTCTATGGCT
TCTGAGGCGG
AAAGAACCAG
CTGGGGCTCG
ATACCGTCGA
CCTCTAGCTA
GAGCTTGGCG
TAATCATGGT
CATAGCTGTT
TCCTGTGTGA
AATTGTTATC
CGCTCACAAT
TCCACACAAC
ATACGAGCCG
GAAGCATAAA
GTGTAAAGCC
TAGGGTGCCT
AATGAGTGAG
CTAACTCACA
TTAATTGCGT
TGCGCTCACT
GCCCGCTTTC
CAGTCGGGAA
ACCTGTCGTG
CCAGCTGCAT
TAATGAATCG
GCCAACGCGC
GGGGAGAGGC
GGTTTGCGTA
TTGGGCGCTC
TTCCGCTTCC
TCGCTCACTG
ACTCGCTGCG
CTCGGTCGTT
CGGCTGCGGC
GAGCGGTATC
AGCTCACTCA
AAGGCGGTAA
TACGGTTATC
CACAGAATCA
GGGGATAACG
CAGGAAAGAA
CATGTGAGCA
AAAGGCCAGC
AAAAGGCCAG
GAACCGTAAA
AAGGCCGCGT
TGCTGGCGTT
TTTCCATAGG
CTCCGCCCCC
CTGACGAGCA
TCACAAAAAT
CGACGCTCAA
GTCAGAGGTG
GCGAAACCCG
ACAGGACTAT
AAAGATACCA
GGCGTTTCCC
CCTGGAAGCT
CCCTCGTGCG
CTCTCCTGTT
CCGACCCTGC
CGCTTACCGG
ATACCTGTCC
GCCTTTCTCC
CTTCGGGAAG
CGTGGCGCTT
TCTCATAGCT
CACGCTGTAG
GTATCTCAGT
TCGGTGTAGG
TCGTTCGCTC
CAAGCTGGGC
TGTGTGCACG
AACCCCCCGT
TCAGCCCGAC
CGCTGCGCCT
TATCCGGTAA
CTATCGTCTT
GAGTCCAACC
CGGTAAGACA
CGACTTATCG
CCACTGGCAG
CAGCCACTGG
TAACAGGATT
AGCAGAGCGA
GGTATGTAGG
CGGTGCTACA
GAGTTCTTGA
AGTGGTGGCC
TAACTACGGC
TACACTAGAA
GAACAGTATT
TGGTATCTGC
GCTCTGCTGA
AGCCAGTTAC
CTTCGGAAAA
AGAGTTGGTA
GCTCTTGATC
CGGCAAACAA
ACCACCGCTG
GTAGCGGTGG
TTTTTTTGTT
TGCAAGCAGC
AGATTACGCG
CAGAAAAAAA
GGATCTCAAG
AAGATCCTTT
GATCTTTTCT
ACGGGGTCTG
ACGCTCAGTG
GAACGAAAAC
TCACGTTAAG
GGATTTTGGT
CATGAGATTA
TCAAAAAGGA
TCTTCACCTA
GATCCTTTTA
AATTAAAAAT
GAAGTTTTAA
ATCAATCTAA
AGTATATATG
AGTAAACTTG
GTCTGACAGT
TACCAATGCT
TAATCAGTGA
GGCACCTATC
TCAGCGATCT
GTCTATTTCG
TTCATCCATA
GTTGCCTGAC
TCCCCGTCGT
GTAGATAACT
ACGATACGGG
AGGGCTTACC
ATCTGGCCCC
AGTGCTGCAA
TGATACCGCG
AGACCCACGC
TCACCGGCTC
CAGATTTATC
AGCAATAAAC
CAGCCAGCCG
GAAGGGCCGA
GCGCAGAAGT
GGTCCTGCAA
CTTTATCCGC
CTCCATCCAG
TCTATTAATT
GTTGCCGGGA
AGCTAGAGTA
AGTAGTTCGC
CAGTTAATAG
TTTGCGCAAC
GTTGTTGCCA
TTGCTACAGG
CATCGTGGTG
TCACGCTCGT
CGTTTGGTAT
GGCTTCATTC
AGCTCCGGTT
CCCAACGATC
AAGGCGAGTT
ACATGATCCC
CCATGTTGTG
CAAAAAAGCG
GTTAGCTCCT
TCGGTCCTCC
GATCGTTGTC
AGAAGTAAGT
TGGCCGCAGT
GTTATCACTC
ATGGTTATGG
CAGCACTGCA
TAATTCTCTT
ACTGTCATGC
CATCCGTAAG
ATGCTTTTCT
GTGACTGGTG
AGTACTCAAC
CAAGTCATTC
TGAGAATAGT
GTATGCGGCG
ACCGAGTTGC
TCTTGCCCGG
CGTCAATACG
GGATAATACC
GCGCCACATA
GCAGAACTTT
AAAAGTGCTC
ATCATTGGAA
AACGTTCTTC
GGGGCGAAAA
CTCTCAAGGA
TCTTACCGCT
GTTGAGATCC
AGTTCGATGT
AACCCACTCG
TGCACCCAAC
TGATCTTCAG
CATCTTTTAC
TTTCACCAGC
GTTTCTGGGT
GAGCAAAAAC
AGGAAGGCAA
AATGCCGCAA
AAAAGGGAAT
AAGGGCGACA
CGGAAATGTT
GAATACTCAT
ACTCTTCCTT
TTTCAATATT
ATTGAAGCAT
TTATCAGGGT
TATTGTCTCA
TGAGCGGATA
CATATTTGAA
TGTATTTAGA
AAAATAAACA
AATAGGGGTT
CCGCGCACAT
TTCCCCGAAA
AGTGCCACCT
GACGTCGACG
GATCGGGAGA
TCGATCTCCC
GATCCCCTAG
GGTCGACTCT
CAGTACAATC
TGCTCTGATG
CCGCATAGTT
AAGCCAGTAT
CTGCTCCCTG
CTTGTGTGTT
G