名称: | pIVT-dCas9-EGFP |
---|---|
别称: | Plasmid#174442 |
基因长: | |
质粒宿主: | 大肠杆菌 |
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: | 绿色 |
启动子: | |
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。
GACGAAAGGG
CCTCGTGATA
CGCCTATTTT
TATAGGTTAA
TGTCATGATA
ATAATGGTTT
CTTAGACGTC
AGGTGGCACT
TTTCGGGGAA
ATGTGCGCGG
AACCCCTATT
TGTTTATTTT
TCTAAATACA
TTCAAATATG
TATCCGCTCA
TGAGACAATA
ACCCTGATAA
ATGCTTCAAT
AATATTGAAA
AAGGAAGAGT
ATGAGTATTC
AACATTTCCG
TGTCGCCCTT
ATTCCCTTTT
TTGCGGCATT
TTGCCTTCCT
GTTTTTGCTC
ACCCAGAAAC
GCTGGTGAAA
GTAAAAGATG
CTGAAGATCA
GTTGGGTGCA
CGAGTGGGTT
ACATCGAACT
GGATCTCAAC
AGCGGTAAGA
TCCTTGAGAG
TTTTCGCCCC
GAAGAACGTT
TTCCAATGAT
GAGCACTTTT
AAAGTTCTGC
TATGTGGCGC
GGTATTATCC
CGTATTGACG
CCGGGCAAGA
GCAACTCGGT
CGCCGCATAC
ACTATTCTCA
GAATGACTTG
GTTGAGTACT
CACCAGTCAC
AGAAAAGCAT
CTTACGGATG
GCATGACAGT
AAGAGAATTA
TGCAGTGCTG
CCATAACCAT
GAGTGATAAC
ACTGCGGCCA
ACTTACTTCT
GACAACGATC
GGAGGACCGA
AGGAGCTAAC
CGCTTTTTTG
CACAACATGG
GGGATCATGT
AACTCGCCTT
GATCGTTGGG
AACCGGAGCT
GAATGAAGCC
ATACCAAACG
ACGAGCGTGA
CACCACGATG
CCTGTAGCAA
TGGCAACAAC
GTTGCGCAAA
CTATTAACTG
GCGAACTACT
TACTCTAGCT
TCCCGGCAAC
AATTAATAGA
CTGGATGGAG
GCGGATAAAG
TTGCAGGACC
ACTTCTGCGC
TCGGCCCTTC
CGGCTGGCTG
GTTTATTGCT
GATAAATCTG
GAGCCGGTGA
GCGTGGGTCT
CGCGGTATCA
TTGCAGCACT
GGGGCCAGAT
GGTAAGCCCT
CCCGTATCGT
AGTTATCTAC
ACGACGGGGA
GTCAGGCAAC
TATGGATGAA
CGAAATAGAC
AGATCGCTGA
GATAGGTGCC
TCACTGATTA
AGCATTGGTA
ACTGTCAGAC
CAAGTTTACT
CATATATACT
TTAGATTGAT
TTAAAACTTC
ATTTTTAATT
TAAAAGGATC
TAGGTGAAGA
TCCTTTTTGA
TAATCTCATG
ACCAAAATCC
CTTAACGTGA
GTTTTCGTTC
CACTGAGCGT
CAGACCCCGT
AGAAAAGATC
AAAGGATCTT
CTTGAGATCC
TTTTTTTCTG
CGCGTAATCT
GCTGCTTGCA
AACAAAAAAA
CCACCGCTAC
CAGCGGTGGT
TTGTTTGCCG
GATCAAGAGC
TACCAACTCT
TTTTCCGAAG
GTAACTGGCT
TCAGCAGAGC
GCAGATACCA
AATACTGTTC
TTCTAGTGTA
GCCGTAGTTA
GGCCACCACT
TCAAGAACTC
TGTAGCACCG
CCTACATACC
TCGCTCTGCT
AATCCTGTTA
CCAGTGGCTG
CTGCCAGTGG
CGATAAGTCG
TGTCTTACCG
GGTTGGACTC
AAGACGATAG
TTACCGGATA
AGGCGCAGCG
GTCGGGCTGA
ACGGGGGGTT
CGTGCACACA
GCCCAGCTTG
GAGCGAACGA
CCTACACCGA
ACTGAGATAC
CTACAGCGTG
AGCTATGAGA
AAGCGCCACG
CTTCCCGAAG
GGAGAAAGGC
GGACAGGTAT
CCGGTAAGCG
GCAGGGTCGG
AACAGGAGAG
CGCACGAGGG
AGCTTCCAGG
GGGAAACGCC
TGGTATCTTT
ATAGTCCTAT
CGGGTTTCGC
CACCTCTGAC
TTGAGCGTCG
ATTTTTGTGA
TGCTCGTCAG
GGGGGCGGAG
CCTATGGAAA
AACGCCAGCA
ACGCGGCCTT
TTTACGGTTC
CTGGCCTTTT
GCTGGCCTTT
TGCTCACATG
TTCTTTCCTG
CGTTATCCCC
TGATTCTGTG
GATAACCGTA
TTACCGCCTT
TGAGTGAGCT
GATACCGCTC
GCCGCAGCCG
AACGACCGAG
CGCAGCGAGT
CAGTGAGCGA
GGAAGCGGAA
GAGCGCCCAA
TACGCAAACC
GCCTCTCCCC
GCGCGTTGGC
CGATTCATTA
ATGCAGCTGG
CACGACAGGT
TTCCCGACTG
GAAAGCGGGC
AGTGAGCGCA
ACGCAATTAA
TGTGAGTTAG
CTCACTCATT
AGGCACCCCA
GGCTTTACAC
TTTATGCTTC
CGGCTCGTAT
GTTGTGTGGA
ATTGTGAGCG
GATAACAATT
TCACACAGGA
AACAGCTATG
ACCATGATTA
CGCCAAGCTC
CAACGATTTA
GGTGACACTA
TAGAAGAGAA
GGAATTAATA
CGACTCACTA
TAGGGAGAGA
GAGAGAATTA
CCCTCACTAA
AGGGAGGAGA
AGCTTGCTTG
TTCTTTTTGC
AGAAGCTCAG
AATAAACGCT
CAACTTTGGC
AGATCAAGCT
TGCATGCCTG
CAGGTCGACT
ATAGGGAGAC
CCAAGCTTGG
CGCGCCGCCA
CCATGGACAA
GAAGTACTCC
ATTGGGCTCG
CTATCGGCAC
AAACAGCGTC
GGCTGGGCCG
TCATTACGGA
CGAGTACAAG
GTGCCGAGCA
AAAAATTCAA
AGTTCTGGGC
AATACCGATC
GCCACAGCAT
AAAGAAGAAC
CTCATTGGCG
CCCTCCTGTT
CGACTCCGGG
GAGACGGCCG
AAGCCACGCG
GCTCAAAAGA
ACAGCACGGC
GCAGATATAC
CCGCAGAAAG
AATCGGATCT
GCTACCTGCA
GGAGATCTTT
AGTAATGAGA
TGGCTAAGGT
GGATGACTCT
TTCTTCCATA
GGCTGGAGGA
GTCCTTTTTG
GTGGAGGAGG
ATAAAAAGCA
CGAGCGCCAC
CCAATCTTTG
GCAATATCGT
GGACGAGGTG
GCGTACCATG
AAAAGTACCC
AACCATATAT
CATCTGAGGA
AGAAGCTTGT
AGACAGTACT
GATAAGGCTG
ACTTGCGGTT
GATCTATCTC
GCGCTGGCGC
ATATGATCAA
ATTTCGGGGA
CACTTCCTCA
TCGAGGGGGA
CCTGAACCCA
GACAACAGCG
ATGTCGACAA
ACTCTTTATC
CAACTGGTTC
AGACTTACAA
TCAGCTTTTC
GAAGAGAACC
CGATCAACGC
ATCCGGAGTT
GACGCCAAAG
CAATCCTGAG
CGCTAGGCTG
TCCAAATCCC
GGCGGCTCGA
AAACCTCATC
GCACAGCTCC
CTGGGGAGAA
GAAGAACGGC
CTGTTTGGTA
ATCTTATCGC
CCTGTCACTC
GGGCTGACCC
CCAACTTTAA
ATCTAACTTC
GACCTGGCCG
AAGATGCCAA
GCTTCAACTG
AGCAAAGACA
CCTACGATGA
TGATCTCGAC
AATCTGCTGG
CCCAGATCGG
CGACCAGTAC
GCAGACCTTT
TTTTGGCGGC
AAAGAACCTG
TCAGACGCCA
TTCTGCTGAG
TGATATTCTG
CGAGTGAACA
CGGAGATCAC
CAAAGCTCCG
CTGAGCGCTA
GTATGATCAA
GCGCTATGAT
GAGCACCACC
AAGACTTGAC
TTTGCTGAAG
GCCCTTGTCA
GACAGCAACT
GCCTGAGAAG
TACAAGGAAA
TTTTCTTCGA
TCAGTCTAAA
AATGGCTACG
CCGGATACAT
TGACGGCGGA
GCAAGCCAGG
AGGAATTTTA
CAAATTTATT
AAGCCCATCT
TGGAAAAAAT
GGACGGCACC
GAGGAGCTGC
TGGTAAAGCT
TAACAGAGAA
GATCTGTTGC
GCAAACAGCG
CACTTTCGAC
AATGGAAGCA
TCCCCCACCA
GATTCACCTG
GGCGAACTGC
ACGCTATCCT
CAGGCGGCAA
GAGGATTTCT
ACCCCTTTTT
GAAAGATAAC
AGGGAAAAGA
TTGAGAAAAT
CCTCACATTT
CGGATACCCT
ACTATGTAGG
CCCCCTCGCC
CGGGGAAATT
CCAGATTCGC
GTGGATGACT
CGCAAATCAG
AAGAGACCAT
CACTCCCTGG
AACTTCGAGG
AAGTCGTGGA
TAAGGGGGCC
TCTGCCCAGT
CCTTCATCGA
AAGGATGACT
AACTTTGATA
AAAATCTGCC
TAACGAAAAG
GTGCTTCCTA
AACACTCTCT
GCTGTACGAG
TACTTCACAG
TTTATAACGA
GCTCACCAAG
GTCAAATACG
TCACAGAAGG
GATGAGAAAG
CCAGCATTCC
TGTCTGGAGA
GCAGAAGAAA
GCTATCGTGG
ACCTCCTCTT
CAAGACGAAC
CGGAAAGTTA
CCGTGAAACA
GCTCAAAGAA
GACTATTTCA
AAAAGATTGA
ATGTTTCGAC
TCTGTTGAAA
TCAGCGGAGT
GGAGGATCGC
TTCAACGCAT
CCCTGGGAAC
GTATCACGAT
CTCCTGAAAA
TCATTAAAGA
CAAGGACTTC
CTGGACAATG
AGGAGAACGA
GGACATTCTT
GAGGACATTG
TCCTCACCCT
TACGTTGTTT
GAAGATAGGG
AGATGATTGA
AGAACGCTTG
AAAACTTACG
CTCATCTCTT
CGACGACAAA
GTCATGAAAC
AGCTCAAGAG
GCGCCGATAT
ACAGGATGGG
GGCGGCTGTC
AAGAAAACTG
ATCAATGGGA
TCCGAGACAA
GCAGAGTGGA
AAGACAATCC
TGGATTTTCT
TAAGTCCGAT
GGATTTGCCA
ACAGGAACTT
CATGCAGTTG
ATCCATGATG
ACTCTCTCAC
CTTTAAGGAG
GACATCCAGA
AAGCACAAGT
TTCTGGCCAG
GGGGACAGTC
TTCACGAGCA
CATCGCTAAT
CTTGCAGGTA
GCCCAGCTAT
CAAAAAGGGA
ATACTGCAGA
CCGTTAAGGT
CGTGGATGAA
CTCGTCAAAG
TAATGGGAAG
GCATAAGCCC
GAGAATATCG
TTATCGAGAT
GGCCCGAGAG
AACCAAACTA
CCCAGAAGGG
ACAGAAGAAC
AGTAGGGAAA
GGATGAAGAG
GATTGAAGAG
GGTATAAAAG
AACTGGGGTC
CCAAATCCTT
AAGGAACACC
CAGTTGAAAA
CACCCAGCTT
CAGAATGAGA
AGCTCTACCT
GTACTACCTG
CAGAACGGCA
GGGACATGTA
CGTGGATCAG
GAACTGGACA
TCAATCGGCT
CTCCGACTAC
GACGTGGATG
CTATCGTGCC
CCAGTCTTTT
CTCAAAGATG
ATTCTATTGA
TAATAAAGTG
TTGACAAGAT
CCGATAAAAA
TAGAGGGAAG
AGTGATAACG
TCCCCTCAGA
AGAAGTTGTC
AAGAAAATGA
AAAATTATTG
GCGGCAGCTG
CTGAACGCCA
AACTGATCAC
ACAACGGAAG
TTCGATAATC
TGACTAAGGC
TGAACGAGGT
GGCCTGTCTG
AGTTGGATAA
AGCCGGCTTC
ATCAAAAGGC
AGCTTGTTGA
GACACGCCAG
ATCACCAAGC
ACGTGGCCCA
AATTCTCGAT
TCACGCATGA
ACACCAAGTA
CGATGAAAAT
GACAAACTGA
TTCGAGAGGT
GAAAGTTATT
ACTCTGAAGT
CTAAGCTGGT
CTCAGATTTC
AGAAAGGACT
TTCAGTTTTA
TAAGGTGAGA
GAGATCAACA
ATTACCACCA
TGCGCATGAT
GCCTACCTGA
ATGCAGTGGT
AGGCACTGCA
CTTATCAAAA
AATATCCCAA
GCTTGAATCT
GAATTTGTTT
ACGGAGACTA
TAAAGTGTAC
GATGTTAGGA
AAATGATCGC
AAAGTCTGAG
CAGGAAATAG
GCAAGGCCAC
CGCTAAGTAC
TTCTTTTACA
GCAATATTAT
GAATTTTTTC
AAGACCGAGA
TTACACTGGC
CAATGGAGAG
ATTCGGAAGC
GACCACTTAT
CGAAACAAAC
GGAGAAACAG
GAGAAATCGT
GTGGGACAAG
GGTAGGGATT
TCGCGACAGT
CCGGAAGGTC
CTGTCCATGC
CGCAGGTGAA
CATCGTTAAA
AAGACCGAAG
TACAGACCGG
AGGCTTCTCC
AAGGAAAGTA
TCCTCCCGAA
AAGGAACAGC
GACAAGCTGA
TCGCACGCAA
AAAAGATTGG
GACCCCAAGA
AATACGGCGG
ATTCGATTCT
CCTACAGTCG
CTTACAGTGT
ACTGGTTGTG
GCCAAAGTGG
AGAAAGGGAA
GTCTAAAAAA
CTCAAAAGCG
TCAAGGAACT
GCTGGGCATC
ACAATCATGG
AGCGATCAAG
CTTCGAAAAA
AACCCCATCG
ACTTTCTCGA
GGCGAAAGGA
TATAAAGAGG
TCAAAAAAGA
CCTCATCATT
AAGCTTCCCA
AGTACTCTCT
CTTTGAGCTT
GAAAACGGCC
GGAAACGAAT
GCTCGCTAGT
GCGGGCGAGC
TGCAGAAAGG
TAACGAGCTG
GCACTGCCCT
CTAAATACGT
TAATTTCTTG
TATCTGGCCA
GCCACTATGA
AAAGCTCAAA
GGGTCTCCCG
AAGATAATGA
GCAGAAGCAG
CTGTTCGTGG
AACAACACAA
ACACTACCTT
GATGAGATCA
TCGAGCAAAT
AAGCGAATTC
TCCAAAAGAG
TGATCCTCGC
CGACGCTAAC
CTCGATAAGG
TGCTTTCTGC
TTACAATAAG
CACAGGGATA
AGCCCATCAG
GGAGCAGGCA
GAAAACATTA
TCCACTTGTT
TACTCTGACC
AACTTGGGCG
CGCCTGCAGC
CTTCAAGTAC
TTCGACACCA
CCATAGACAG
AAAGCGGTAC
ACCTCTACAA
AGGAGGTCCT
GGACGCCACA
CTGATTCATC
AGTCAATTAC
GGGGCTCTAT
GAAACAAGAA
TCGACCTCTC
TCAGCTCGGT
GGAGACGCTA
GCGGCTCTAG
AATGGTGAGC
AAGGGCGAGG
AGCTGTTCAC
CGGGGTGGTG
CCCATCCTGG
TCGAGCTGGA
CGGCGACGTA
AACGGCCACA
AGTTCAGCGT
GTCCGGCGAG
GGCGAGGGCG
ATGCCACCTA
CGGCAAGCTG
ACCCTGAAGT
TCATCTGCAC
CACCGGCAAG
CTGCCCGTGC
CCTGGCCCAC
CCTCGTGACC
ACCCTGACCT
ACGGCGTGCA
GTGCTTCAGC
CGCTACCCCG
ACCACATGAA
GCAGCACGAC
TTCTTCAAGT
CCGCCATGCC
CGAAGGCTAC
GTCCAGGAGC
GCACCATCTT
CTTCAAGGAC
GACGGCAACT
ACAAGACCCG
CGCCGAGGTG
AAGTTCGAGG
GCGACACCCT
GGTGAACCGC
ATCGAGCTGA
AGGGCATCGA
CTTCAAGGAG
GACGGCAACA
TCCTGGGGCA
CAAGCTGGAG
TACAACTACA
ACAGCCACAA
CGTCTATATC
ATGGCCGACA
AGCAGAAGAA
CGGCATCAAG
GTGAACTTCA
AGATCCGCCA
CAACATCGAG
GACGGCAGCG
TGCAGCTCGC
CGACCACTAC
CAGCAGAACA
CCCCCATCGG
CGACGGCCCC
GTGCTGCTGC
CCGACAACCA
CTACCTGAGC
ACCCAGTCCG
CCCTGAGCAA
AGACCCCAAC
GAGAAGCGCG
ATCACATGGT
CCTGCTGGAG
TTCGTGACCG
CCGCCGGGAT
CACTCTCGGC
ATGGACGAGC
TGTACAAGTA
GTGACTGACT
AGGATCTGGT
TACCACTAAA
CCAGCCTCAA
GAACACCCGA
ATGGAGTCTC
TAAGCTACAT
AATACCAACT
TACACTTTAC
AAAATGTTGT
CCCCCAAAAT
GTAGCCATTC
GTATCTGCTC
CTAATAAAAA
GAAAGTTTCT
TCACATTCTA
AAAAAAAAAA
AAAAAAAAAA
AAAAAAAAAA
AnCCCCCCCC
CCCCAAGGGC
GAATTCACTG
GCCGTCGTTT
TACAACGTCG
TGACTGGGAA
AACCCTGGCG
TTACCCAACT
TAATCGCCTT
GCAGCACATC
CCCCTTTCGC
CAGCTGGCGT
AATAGCGAAG
AGGCCCGCAC
CGATCGCCCT
TCCCAACAGT
TGCGCAGCCT
GAATGGCGAA
TGGCGCCTGA
TGCGGTATTT
TCTCCTTACG
CATCTGTGCG
GTATTTCACA
CCGCATATGG
TGCACTCTCA
GTACAATCTG
CTCTGATGCC
GCATAGTTAA
GCCAGCCCCG
ACACCCGCCA
ACACCCGCTG
ACGCGCCCTG
ACGGGCTTGT
CTGCTCCCGG
CATCCGCTTA
CAGACAAGCT
GTGACCGTCT
CCGGGAGCTG
CATGTGTCAG
AGGTTTTCAC
CGTCATCACC
GAAACGCGCG
A