名称: | pX853-U6-gRNA-CBh-SpCas9 |
---|---|
别称: | Plasmid#62885 |
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: | |
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
ATGCTAGATT
TAGCTAGCTT
AATCCTCGAC
AAGAAGTACA
GCATCGGCCT
GGACATCGGC
ACCAACTCTG
TGGGCTGGGC
CGTGATCACC
GACGAGTACA
AGGTGCCCAG
CAAGAAATTC
AAGGTGCTGG
GCAACACCGA
CCGGCACAGC
ATCAAGAAGA
ACCTGATCGG
AGCCCTGCTG
TTCGACAGCG
GCGAAACAGC
CGAGGCCACC
CGGCTGAAGA
GAACCGCCAG
AAGAAGATAC
ACCAGACGGA
AGAACCGGAT
CTGCTATCTG
CAAGAGATCT
TCAGCAACGA
GATGGCCAAG
GTGGACGACA
GCTTCTTCCA
CAGACTGGAA
GAGTCCTTCC
TGGTGGAAGA
GGATAAGAAG
CACGAGCGGC
ACCCCATCTT
CGGCAACATC
GTGGACGAGG
TGGCCTACCA
CGAGAAGTAC
CCCACCATCT
ACCACCTGAG
AAAGAAACTG
GTGGACAGCA
CCGACAAGGC
CGACCTGCGG
CTGATCTATC
TGGCCCTGGC
CCACATGATC
AAGTTCCGGG
GCCACTTCCT
GATCGAGGGC
GACCTGAACC
CCGACAACAG
CGACGTGGAC
AAGCTGTTCA
TCCAGCTGGT
GCAGACCTAC
AACCAGCTGT
TCGAGGAAAA
CCCCATCAAC
GCCAGCGGCG
TGGACGCCAA
GGCCATCCTG
TCTGCCAGAC
TGAGCAAGAG
CAGACGGCTG
GAAAATCTGA
TCGCCCAGCT
GCCCGGCGAG
AAGAAGAATG
GCCTGTTCGG
AAACCTGATT
GCCCTGAGCC
TGGGCCTGAC
CCCCAACTTC
AAGAGCAACT
TCGACCTGGC
CGAGGATGCC
AAACTGCAGC
TGAGCAAGGA
CACCTACGAC
GACGACCTGG
ACAACCTGCT
GGCCCAGATC
GGCGACCAGT
ACGCCGACCT
GTTTCTGGCC
GCCAAGAACC
TGTCCGACGC
CATCCTGCTG
AGCGACATCC
TGAGAGTGAA
CACCGAGATC
ACCAAGGCCC
CCCTGAGCGC
CTCTATGATC
AAGAGATACG
ACGAGCACCA
CCAGGACCTG
ACCCTGCTGA
AAGCTCTCGT
GCGGCAGCAG
CTGCCTGAGA
AGTACAAAGA
GATTTTCTTC
GACCAGAGCA
AGAACGGCTA
CGCCGGCTAC
ATTGACGGCG
GAGCCAGCCA
GGAAGAGTTC
TACAAGTTCA
TCAAGCCCAT
CCTGGAAAAG
ATGGACGGCA
CCGAGGAACT
GCTCGTGAAG
CTGAACAGAG
AGGACCTGCT
GCGGAAGCAG
CGGACCTTCG
ACAACGGCAG
CATCCCCCAC
CAGATCCACC
TGGGAGAGCT
GCACGCCATT
CTGCGGCGGC
AGGAAGATTT
TTACCCATTC
CTGAAGGACA
ACCGGGAAAA
GATCGAGAAG
ATCCTGACCT
TCCGCATCCC
CTACTACGTG
GGCCCTCTGG
CCAGGGGAAA
CAGCAGATTC
GCCTGGATGA
CCAGAAAGAG
CGAGGAAACC
ATCACCCCCT
GGAACTTCGA
GGAAGTGGTG
GACAAGGGCG
CTTCCGCCCA
GAGCTTCATC
GAGCGGATGA
CCAACTTCGA
TAAGAACCTG
CCCAACGAGA
AGGTGCTGCC
CAAGCACAGC
CTGCTGTACG
AGTACTTCAC
CGTGTATAAC
GAGCTGACCA
AAGTGAAATA
CGTGACCGAG
GGAATGAGAA
AGCCCGCCTT
CCTGAGCGGC
GAGCAGAAAA
AGGCCATCGT
GGACCTGCTG
TTCAAGACCA
ACCGGAAAGT
GACCGTGAAG
CAGCTGAAAG
AGGACTACTT
CAAGAAAATC
GAGGGTGGCG
GTGGCTCGGG
TGGCGGTGGC
TCGGGTGGCG
GTGGCTCGGA
GATGTGGCAC
GAGGGCCTGG
AGGAGGCCAG
CAGACTGTAC
TTCGGCGAGA
GAAACGTGAA
GGGCATGTTC
GAGGTGCTGG
AGCCCCTGCA
CGCCATGATG
GAGAGAGGCC
CCCAGACCCT
GAAGGAGACC
AGCTTCAACC
AGGCCTACGG
CAGAGACCTG
ATGGAGGCCC
AGGAGTGGTG
CAGAAAGTAC
ATGAAGAGCG
GCAACGTGAA
GGACCTGACC
CAGGCCTGGG
ACCTGTACTA
CCACGTGTTC
AGAAGAATCA
GCAAGCAGTA
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