名称: | pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000) |
---|---|
别称: | Plasmid #139996 |
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。
AGCATCAAGA
AGAACCTGAT
CGGAGCCCTG
CTGTTCGACA
GCGGCGAAAC
AGCCGAGGCC
ACCCGGCTGA
AGAGAACCGC
CAGAAGAAGA
TACACCAGAC
GGAAGAACCG
GATCTGCTAT
CTGCAAGAGA
TCTTCAGCAA
CGAGATGGCC
AAGGTGGACG
ACAGCTTCTT
CCACAGACTG
GAAGAGTCCT
TCCTGGTGGA
AGAGGATAAG
AAGCACGAGC
GGCACCCCAT
CTTCGGCAAC
ATCGTGGACG
AGGTGGCCTA
CCACGAGAAG
TACCCCACCA
TCTACCACCT
GAGAAAGAAA
CTGGTGGACA
GCACCGACAA
GGCCGACCTG
CGGCTGATCT
ATCTGGCCCT
GGCCCACATG
ATCAAGTTCC
GGGGCCACTT
CCTGATCGAG
GGCGACCTGA
ACCCCGACAA
CAGCGACGTG
GACAAGCTGT
TCATCCAGCT
GGTGCAGACC
TACAACCAGC
TGTTCGAGGA
AAACCCCATC
AACGCCAGCG
GCGTGGACGC
CAAGGCCATC
CTGTCTGCCA
GACTGAGCAA
GAGCAGACGG
CTGGAAAATC
TGATCGCCCA
GCTGCCCGGC
GAGAAGAAGA
ATGGCCTGTT
CGGAAACCTG
ATTGCCCTGA
GCCTGGGCCT
GACCCCCAAC
TTCAAGAGCA
ACTTCGACCT
GGCCGAGGAT
GCCAAACTGC
AGCTGAGCAA
GGACACCTAC
GACGACGACC
TGGACAACCT
GCTGGCCCAG
ATCGGCGACC
AGTACGCCGA
CCTGTTTCTG
GCCGCCAAGA
ACCTGTCCGA
CGCCATCCTG
CTGAGCGACA
TCCTGAGAGT
GAACACCGAG
ATCACCAAGG
CCCCCCTGAG
CGCCTCTATG
ATCAAGAGAT
ACGACGAGCA
CCACCAGGAC
CTGACCCTGC
TGAAAGCTCT
CGTGCGGCAG
CAGCTGCCTG
AGAAGTACAA
AGAGATTTTC
TTCGACCAGA
GCAAGAACGG
CTACGCCGGC
TACATTGACG
GCGGAGCCAG
CCAGGAAGAG
TTCTACAAGT
TCATCAAGCC
CATCCTGGAA
AAGATGGACG
GCACCGAGGA
ACTGCTCGTG
AAGCTGAACA
GAGAGGACCT
GCTGCGGAAG
CAGCGGACCT
TCGACAACGG
CAGCATCCCC
CACCAGATCC
ACCTGGGAGA
GCTGCACGCC
ATTCTGCGGC
GGCAGGAAGA
TTTTTACCCA
TTCCTGAAGG
ACAACCGGGA
AAAGATCGAG
AAGATCCTGA
CCTTCCGCAT
CCCCTACTAC
GTGGGCCCTC
TGGCCAGGGG
AAACAGCAGA
TTCGCCTGGA
TGACCAGAAA
GAGCGAGGAA
ACCATCACCC
CCTGGAACTT
CGAGGAAGTG
GTGGACAAGG
GCGCTTCCGC
CCAGAGCTTC
ATCGAGCGGA
TGACCGCCTT
CGATAAGAAC
CTGCCCAACG
AGAAGGTGCT
GCCCAAGCAC
AGCCTGCTGT
ACGAGTACTT
CACCGTGTAT
AACGAGCTGA
CCAAAGTGAA
ATACGTGACC
GAGGGAATGA
GAAAGCCCGC
CTTCCTGAGC
GGCGAGCAGA
AAAAGGCCAT
CGTGGACCTG
CTGTTCAAGA
CCAACCGGAA
AGTGACCGTG
AAGCAGCTGA
AAGAGGACTA
CTTCAAGAAA
ATCGAGTGCT
TCGACTCCGT
GGAAATCTCC
GGCGTGGAAG
ATCGGTTCAA
CGCCTCCCTG
GGCACATACC
ACGATCTGCT
GAAAATTATC
AAGGACAAGG
ACTTCCTGGA
CAATGAGGAA
AACGAGGACA
TTCTGGAAGA
TATCGTGCTG
ACCCTGACAC
TGTTTGAGGA
CAGAGAGATG
ATCGAGGAAC
GGCTGAAAAC
CTATGCCCAC
CTGTTCGACG
ACAAAGTGAT
GAAGCAGCTG
AAGCGGCGGA
GATACACCGG
CTGGGGCGCC
CTGAGCCGGA
AGCTGATCAA
CGGCATCCGG
GACAAGCAGT
CCGGCAAGAC
AATCCTGGAT
TTCCTGAAGT
CCGACGGCTT
CGCCAACAGA
AACTTCATGG
CCCTGATCCA
CGACGACAGC
CTGACCTTTA
AAGAGGACAT
CCAGAAAGCC
CAGGTGTCCG
GCCAGGGCGA
TAGCCTGCAC
GAGCACATTG
CCAATCTGGC
CGGCAGCCCC
GCCATTAAGA
AGGGCATCCT
GCAGACAGTG
AAGGTGGTGG
ACGAGCTCGT
GAAAGTGATG
GGCCGGCACA
AGCCCGAGAA
CATCGTGATC
GAAATGGCCA
GAGAGAACCA
GACCACCCAG
AAGGGACAGA
AGAACAGCCG
CGAGAGAATG
AAGCGGATCG
AAGAGGGCAT
CAAAGAGCTG
GGCAGCCAGA
TCCTGAAAGA
ACACCCCGTG
GAAAACACCC
AGCTGCAGAA
CGAGAAGCTG
TACCTGTACT
ACCTGCAGAA
TGGGCGGGAT
ATGTACGTGG
ACCAGGAACT
GGACATCAAC
CGGCTGTCCG
ACTACGATGT
GGACCATATC
GTGCCTCAGA
GCTTTCTGAA
GGACGACTCC
ATCGACAACA
AGGTGCTGAC
CAGAAGCGAC
AAGAACCGGG
GCAAGAGCGA
CAACGTGCCC
TCCGAAGAGG
TCGTGAAGAA
GATGAAGAAC
TACTGGCGGC
AGCTGCTGAA
CGCCAAGCTG
ATTACCCAGA
GAAAGTTCGA
CAATCTGACC
AAGGCCGAGA
GAGGCGGCCT
GAGCGAACTG
GATAAGGCCG
GCTTCATCAA
GAGACAGCTG
GTGGAAACCC
GGGCCATCAC
AAAGCACGTG
GCACAGATCC
TGGACTCCCG
GATGAACACT
AAGTACGACG
AGAATGACAA
GCTGATCCGG
GAAGTGAAAG
TGATCACCCT
GAAGTCCAAG
CTGGTGTCCG
ATTTCCGGAA
GGATTTCCAG
TTTTACAAAG
TGCGCGAGAT
CAACAACTAC
CACCACGCCC
ACGACGCCTA
CCTGAACGCC
GTCGTGGGAA
CCGCCCTGAT
CAAAAAGTAC
CCTAAGCTGG
AAAGCGAGTT
CGTGTACGGC
GACTACAAGG
TGTACGACGT
GCGGAAGATG
ATCGCCAAGA
GCGAGCAGGA
AATCGGCAAG
GCTACCGCCA
AGTACTTCTT
CTACAGCAAC
ATCATGAACT
TTTTCAAGAC
CGAGATTACC
CTGGCCAACG
GCGAGATCCG
GAAGCGGCCT
CTGATCGAGA
CAAACGGCGA
AACCGGGGAG
ATCGTGTGGG
ATAAGGGCCG
GGATTTTGCC
ACCGTGCGGA
AAGTGCTGAG
CATGCCCCAA
GTGAATATCG
TGAAAAAGAC
CGAGGTGCAG
ACAGGCGGCT
TCAGCAAAGA
GTCTATCCTG
CCCAAGAGGA
ACAGCGATAA
GCTGATCGCC
AGAAAGAAGG
ACTGGGACCC
TAAGAAGTAC
GGCGGCTTCC
TGTGGCCCAC
CGTGGCCTAT
TCTGTGCTGG
TGGTGGCCAA
AGTGGAAAAG
GGCAAGTCCA
AGAAACTGAA
GAGTGTGAAA
GAGCTGCTGG
GGATCACCAT
CATGGAAAGA
AGCAGCTTCG
AGAAGAATCC
CATCGACTTT
CTGGAAGCCA
AGGGCTACAA
AGAAGTGAAA
AAGGACCTGA
TCATCAAGCT
GCCTAAGTAC
TCCCTGTTCG
AGCTGGAAAA
CGGCCGGAAG
AGAATGCTGG
CCTCTGCCAA
GCAGCTGCAG
AAGGGAAACG
AACTGGCCCT
GCCCTCCAAA
TATGTGAACT
TCCTGTACCT
GGCCAGCCAC
TATGAGAAGC
TGAAGGGCTC
CCCCGAGGAT
AATGAGCAGA
AACAGCTGTT
TGTGGAACAG
CACAAGCACT
ACCTGGACGA
GATCATCGAG
CAGATCAGCG
AGTTCTCCAA
GAGAGTGATC
CTGGCCGACG
CTAATCTGGA
CAAAGTGCTG
TCCGCCTACA
ACAAGCACCG
GGATAAGCCC
ATCAGAGAGC
AGGCCGAGAA
TATCATCCAC
CTGTTTACCC
TGACCAATCT
GGGAGCCCCT
GCCGCCTTCA
AGTACTTTGA
CACCACCATC
GACCGGAAGC
AGTACAGAAG
CACCAAAGAG
GTGCTGGACG
CCACCCTGAT
CCACCAGAGC
ATCACCGGCC
TGTACGAGAC
ACGGATCGAC
CTGTCTCAGC
TGGGAGGCGA
CGGATCCGGC
GGAGGCGGAA
GCGGGAAAAG
AACCGCCGAC
GGCAGCGAAT
TCGAGCCCAA
GAAGAAGAGG
AAAGTCTCGA
GCGGAGGCGA
CTACAAAGAC
CATGACGGTG
ATTATAAAGA
TCATGACATC
GATTACAAGG
ATGACGATGA
CAAGGGAGGC
GGTGGAAGCG
GCGCAACAAA
CTTCTCTCTG
CTGAAACAAG
CCGGAGATGT
CGAAGAGAAT
CCTGGACCGA
TGGTGAGCAA
GGGCGAGGAG
CTGTTCACCG
GGGTGGTGCC
CATCCTGGTC
GAGCTGGACG
GCGACGTAAA
CGGCCACAAG
TTCAGCGTGT
CCGGCGAGGG
CGAGGGCGAT
GCCACCTACG
GCAAGCTGAC
CCTGAAGTTC
ATCTGCACCA
CCGGCAAGCT
GCCCGTGCCC
TGGCCCACCC
TCGTGACCAC
CCTGACCTAC
GGCGTGCAGT
GCTTCAGCCG
CTACCCCGAC
CACATGAAGC
AGCACGACTT
CTTCAAGTCC
GCCATGCCCG
AAGGCTACGT
CCAGGAGCGC
ACCATCTTCT
TCAAGGACGA
CGGCAACTAC
AAGACCCGCG
CCGAGGTGAA
GTTCGAGGGC
GACACCCTGG
TGAACCGCAT
CGAGCTGAAG
GGCATCGACT
TCAAGGAGGA
CGGCAACATC
CTGGGGCACA
AGCTGGAGTA
CAACTACAAC
AGCCACAACG
TCTATATCAT
GGCCGACAAG
CAGAAGAACG
GCATCAAGGT
GAACTTCAAG
ATCCGCCACA
ACATCGAGGA
CGGCAGCGTG
CAGCTCGCCG
ACCACTACCA
GCAGAACACC
CCCATCGGCG
ACGGCCCCGT
GCTGCTGCCC
GACAACCACT
ACCTGAGCAC
CCAGTCCGCC
CTGAGCAAAG
ACCCCAACGA
GAAGCGCGAT
CACATGGTCC
TGCTGGAGTT
CGTGACCGCC
GCCGGGATCA
CTCTCGGCAT
GGACGAGCTG
TACAAGTGAC
CGGTCATCAT
CACCATCACC
ATTGAGTTTA
AACCCGCTGA
TCAGCCTCGA
CTGTGCCTTC
TAGTTGCCAG
CCATCTGTTG
TTTGCCCCTC
CCCCGTGCCT
TCCTTGACCC
TGGAAGGTGC
CACTCCCACT
GTCCTTTCCT
AATAAAATGA
GGAAATTGCA
TCGCATTGTC
TGAGTAGGTG
TCATTCTATT
CTGGGGGGTG
GGGTGGGGCA
GGACAGCAAG
GGGGAGGATT
GGGAAGACAA
TAGCAGGCAT
GCTGGGGATG
CGGTGGGCTC
TATGGCTTCT
GAGGCGGAAA
GAACCAGCTG
GGGCTCGATA
CCGTCGACCT
CTAGCTAGAG
CTTGGCGTAA
TCATGGTCAT
AGCTGTTTCC
TGTGTGAAAT
TGTTATCCGC
TCACAATTCC
ACACAACATA
CGAGCCGGAA
GCATAAAGTG
TAAAGCCTAG
GGTGCCTAAT
GAGTGAGCTA
ACTCACATTA
ATTGCGTTGC
GCTCACTGCC
CGCTTTCCAG
TCGGGAAACC
TGTCGTGCCA
GCTGCATTAA
TGAATCGGCC
AACGCGCGGG
GAGAGGCGGT
TTGCGTATTG
GGCGCTCTTC
CGCTTCCTCG
CTCACTGACT
CGCTGCGCTC
GGTCGTTCGG
CTGCGGCGAG
CGGTATCAGC
TCACTCAAAG
GCGGTAATAC
GGTTATCCAC
AGAATCAGGG
GATAACGCAG
GAAAGAACAT
GTGAGCAAAA
GGCCAGCAAA
AGGCCAGGAA
CCGTAAAAAG
GCCGCGTTGC
TGGCGTTTTT
CCATAGGCTC
CGCCCCCCTG
ACGAGCATCA
CAAAAATCGA
CGCTCAAGTC
AGAGGTGGCG
AAACCCGACA
GGACTATAAA
GATACCAGGC
GTTTCCCCCT
GGAAGCTCCC
TCGTGCGCTC
TCCTGTTCCG
ACCCTGCCGC
TTACCGGATA
CCTGTCCGCC
TTTCTCCCTT
CGGGAAGCGT
GGCGCTTTCT
CATAGCTCAC
GCTGTAGGTA
TCTCAGTTCG
GTGTAGGTCG
TTCGCTCCAA
GCTGGGCTGT
GTGCACGAAC
CCCCCGTTCA
GCCCGACCGC
TGCGCCTTAT
CCGGTAACTA
TCGTCTTGAG
TCCAACCCGG
TAAGACACGA
CTTATCGCCA
CTGGCAGCAG
CCACTGGTAA
CAGGATTAGC
AGAGCGAGGT
ATGTAGGCGG
TGCTACAGAG
TTCTTGAAGT
GGTGGCCTAA
CTACGGCTAC
ACTAGAAGAA
CAGTATTTGG
TATCTGCGCT
CTGCTGAAGC
CAGTTACCTT
CGGAAAAAGA
GTTGGTAGCT
CTTGATCCGG
CAAACAAACC
ACCGCTGGTA
GCGGTGGTTT
TTTTGTTTGC
AAGCAGCAGA
TTACGCGCAG
AAAAAAAGGA
TCTCAAGAAG
ATCCTTTGAT
CTTTTCTACG
GGGTCTGACG
CTCAGTGGAA
CGAAAACTCA
CGTTAAGGGA
TTTTGGTCAT
GAGATTATCA
AAAAGGATCT
TCACCTAGAT
CCTTTTAAAT
TAAAAATGAA
GTTTTAAATC
AATCTAAAGT
ATATATGAGT
AAACTTGGTC
TGACAGTTAC
CAATGCTTAA
TCAGTGAGGC
ACCTATCTCA
GCGATCTGTC
TATTTCGTTC
ATCCATAGTT
GCCTGACTCC
CCGTCGTGTA
GATAACTACG
ATACGGGAGG
GCTTACCATC
TGGCCCCAGT
GCTGCAATGA
TACCGCGAGA
CCCACGCTCA
CCGGCTCCAG
ATTTATCAGC
AATAAACCAG
CCAGCCGGAA
GGGCCGAGCG
CAGAAGTGGT
CCTGCAACTT
TATCCGCCTC
CATCCAGTCT
ATTAATTGTT
GCCGGGAAGC
TAGAGTAAGT
AGTTCGCCAG
TTAATAGTTT
GCGCAACGTT
GTTGCCATTG
CTACAGGCAT
CGTGGTGTCA
CGCTCGTCGT
TTGGTATGGC
TTCATTCAGC
TCCGGTTCCC
AACGATCAAG
GCGAGTTACA
TGATCCCCCA
TGTTGTGCAA
AAAAGCGGTT
AGCTCCTTCG
GTCCTCCGAT
CGTTGTCAGA
AGTAAGTTGG
CCGCAGTGTT
ATCACTCATG
GTTATGGCAG
CACTGCATAA
TTCTCTTACT
GTCATGCCAT
CCGTAAGATG
CTTTTCTGTG
ACTGGTGAGT
ACTCAACCAA
GTCATTCTGA
GAATAGTGTA
TGCGGCGACC
GAGTTGCTCT
TGCCCGGCGT
CAATACGGGA
TAATACCGCG
CCACATAGCA
GAACTTTAAA
AGTGCTCATC
ATTGGAAAAC
GTTCTTCGGG
GCGAAAACTC
TCAAGGATCT
TACCGCTGTT
GAGATCCAGT
TCGATGTAAC
CCACTCGTGC
ACCCAACTGA
TCTTCAGCAT
CTTTTACTTT
CACCAGCGTT
TCTGGGTGAG
CAAAAACAGG
AAGGCAAAAT
GCCGCAAAAA
AGGGAATAAG
GGCGACACGG
AAATGTTGAA
TACTCATACT
CTTCCTTTTT
CAATATTATT
GAAGCATTTA
TCAGGGTTAT
TGTCTCATGA
GCGGATACAT
ATTTGAATGT
ATTTAGAAAA
ATAAACAAAT
AGGGGTTCCG
CGCACATTTC
CCCGAAAAGT
GCCACCTGAC
GTCGACGGAT
CGGGAGATCG
ATCTCCCGAT
CCCCTAGGGT
CGACTCTCAG
TACAATCTGC
TCTGATGCCG
CATAGTTAAG
CCAGTATCTG
CTCCCTGCTT
GTGTGTTGGA
GGTCGCTGAG
TAGTGCGCGA
GCAAAATTTA
AGCTACAACA
AGGCAAGGCT
TGACCGACAA
TTGCATGAAG
AATCTGCTTA
GGGTTAGGCG
TTTTGCGCTG
CTTCGCGATG
TACGGGCCAG
ATATACGCGT
TGACATTGAT
TATTGACTAG
TTATTAATAG
TAATCAATTA
CGGGGTCATT
AGTTCATAGC
CCATATATGG
AGTTCCGCGT
TACATAACTT
ACGGTAAATG
GCCCGCCTGG
CTGACCGCCC
AACGACCCCC
GCCCATTGAC
GTCAATAATG
ACGTATGTTC
CCATAGTAAC
GCCAATAGGG
ACTTTCCATT
GACGTCAATG
GGTGGAGTAT
TTACGGTAAA
CTGCCCACTT
GGCAGTACAT
CAAGTGTATC
ATATGCCAAG
TACGCCCCCT
ATTGACGTCA
ATGACGGTAA
ATGGCCCGCC
TGGCATTATG
CCCAGTACAT
GACCTTATGG
GACTTTCCTA
CTTGGCAGTA
CATCTACGTA
TTAGTCATCG
CTATTACCAT
GGTGATGCGG
TTTTGGCAGT
ACATCAATGG
GCGTGGATAG
CGGTTTGACT
CACGGGGATT
TCCAAGTCTC
CACCCCATTG
ACGTCAATGG
GAGTTTGTTT
TGGCACCAAA
ATCAACGGGA
CTTTCCAAAA
TGTCGTAACA
ACTCCGCCCC
ATTGACGCAA
ATGGGCGGTA
GGCGTGTACG
GTGGGAGGTC
TATATAAGCA
GAGCTGGTTT
AGTGAACCGT
CAGATCCGCT
AGAGATCCGC
GGCCGCTAAT
ACGACTCACT
ATAGGGAGAG
CCGCCACCAT
GGACAAGAAG
TACAGCATCG
GCCTGGACAT
CGGCACCAAC
TCTGTGGGCT
GGGCCGTGAT
CACCGACGAG
TACAAGGTGC
CCAGCAAGAA
ATTCAAGGTG
CTGGGCAACA
CCGACCGGCA
C