名称: | pX854-U6-gRNA-CBh-SpCas9 |
---|---|
别称: | Plasmid#62886 |
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
ATGGCCCCAA
AGAAGAAGCG
GAAGGTCGGT
ATCCACGGAG
TCCCAGCAGC
CGGCGTGCAG
GTGGAGACCA
TCAGCCCCGG
CGACGGCAGA
ACCTTCCCCA
AGAGAGGCCA
GACCTGCGTG
GTGCACTACA
CCGGCATGCT
GGAGGACGGC
AAGAAGTTCG
ACAGCAGCAG
AGACAGAAAC
AAGCCCTTCA
AGTTCATGCT
GGGCAAGCAG
GAGGTGATCA
GAGGCTGGGA
GGAGGGCGTG
GCCCAGATGA
GCGTGGGCCA
GAGAGCCAAG
CTGACCATCA
GCCCCGACTA
CGCCTACGGC
GCCACCGGCC
ACCCCGGCAT
CATCCCTCCT
CACGCCACCC
TGGTGTTCGA
CGTGGAGCTG
CTGAAGCTGG
AGGGTGGCGG
TGGCTCGGGT
GGCGGTGGCT
CGGGTGGCGG
TGGCTCGTGC
TTCGACTCCG
TGGAAATCTC
CGGCGTGGAA
GATCGGTTCA
ACGCCTCCCT
GGGCACATAC
CACGATCTGC
TGAAAATTAT
CAAGGACAAG
GACTTCCTGG
ACAATGAGGA
AAACGAGGAC
ATTCTGGAAG
ATATCGTGCT
GACCCTGACA
CTGTTTGAGG
ACAGAGAGAT
GATCGAGGAA
CGGCTGAAAA
CCTATGCCCA
CCTGTTCGAC
GACAAAGTGA
TGAAGCAGCT
GAAGCGGCGG
AGATACACCG
GCTGGGGCAG
GCTGAGCCGG
AAGCTGATCA
ACGGCATCCG
GGACAAGCAG
TCCGGCAAGA
CAATCCTGGA
TTTCCTGAAG
TCCGACGGCT
TCGCCAACAG
AAACTTCATG
CAGCTGATCC
ACGACGACAG
CCTGACCTTT
AAAGAGGACA
TCCAGAAAGC
CCAGGTGTCC
GGCCAGGGCG
ATAGCCTGCA
CGAGCACATT
GCCAATCTGG
CCGGCAGCCC
CGCCATTAAG
AAGGGCATCC
TGCAGACAGT
GAAGGTGGTG
GACGAGCTCG
TGAAAGTGAT
GGGCCGGCAC
AAGCCCGAGA
ACATCGTGAT
CGAAATGGCC
AGAGAGAACC
AGACCACCCA
GAAGGGACAG
AAGAACAGCC
GCGAGAGAAT
GAAGCGGATC
GAAGAGGGCA
TCAAAGAGCT
GGGCAGCCAG
ATCCTGAAAG
AACACCCCGT
GGAAAACACC
CAGCTGCAGA
ACGAGAAGCT
GTACCTGTAC
TACCTGCAGA
ATGGGCGGGA
TATGTACGTG
GACCAGGAAC
TGGACATCAA
CCGGCTGTCC
GACTACGATG
TGGACCATAT
CGTGCCTCAG
AGCTTTCTGA
AGGACGACTC
CATCGACAAC
AAGGTGCTGA
CCAGAAGCGA
CAAGAACCGG
GGCAAGAGCG
ACAACGTGCC
CTCCGAAGAG
GTCGTGAAGA
AGATGAAGAA
CTACTGGCGG
CAGCTGCTGA
ACGCCAAGCT
GATTACCCAG
AGAAAGTTCG
ACAATCTGAC
CAAGGCCGAG
AGAGGCGGCC
TGAGCGAACT
GGATAAGGCC
GGCTTCATCA
AGAGACAGCT
GGTGGAAACC
CGGCAGATCA
CAAAGCACGT
GGCACAGATC
CTGGACTCCC
GGATGAACAC
TAAGTACGAC
GAGAATGACA
AGCTGATCCG
GGAAGTGAAA
GTGATCACCC
TGAAGTCCAA
GCTGGTGTCC
GATTTCCGGA
AGGATTTCCA
GTTTTACAAA
GTGCGCGAGA
TCAACAACTA
CCACCACGCC
CACGACGCCT
ACCTGAACGC
CGTCGTGGGA
ACCGCCCTGA
TCAAAAAGTA
CCCTAAGCTG
GAAAGCGAGT
TCGTGTACGG
CGACTACAAG
GTGTACGACG
TGCGGAAGAT
GATCGCCAAG
AGCGAGCAGG
AAATCGGCAA
GGCTACCGCC
AAGTACTTCT
TCTACAGCAA
CATCATGAAC
TTTTTCAAGA
CCGAGATTAC
CCTGGCCAAC
GGCGAGATCC
GGAAGCGGCC
TCTGATCGAG
ACAAACGGCG
AAACCGGGGA
GATCGTGTGG
GATAAGGGCC
GGGATTTTGC
CACCGTGCGG
AAAGTGCTGA
GCATGCCCCA
AGTGAATATC
GTGAAAAAGA
CCGAGGTGCA
GACAGGCGGC
TTCAGCAAAG
AGTCTATCCT
GCCCAAGAGG
AACAGCGATA
AGCTGATCGC
CAGAAAGAAG
GACTGGGACC
CTAAGAAGTA
CGGCGGCTTC
GACAGCCCCA
CCGTGGCCTA
TTCTGTGCTG
GTGGTGGCCA
AAGTGGAAAA
GGGCAAGTCC
AAGAAACTGA
AGAGTGTGAA
AGAGCTGCTG
GGGATCACCA
TCATGGAAAG
AAGCAGCTTC
GAGAAGAATC
CCATCGACTT
TCTGGAAGCC
AAGGGCTACA
AAGAAGTGAA
AAAGGACCTG
ATCATCAAGC
TGCCTAAGTA
CTCCCTGTTC
GAGCTGGAAA
ACGGCCGGAA
GAGAATGCTG
GCCTCTGCCG
GCGAACTGCA
GAAGGGAAAC
GAACTGGCCC
TGCCCTCCAA
ATATGTGAAC
TTCCTGTACC
TGGCCAGCCA
CTATGAGAAG
CTGAAGGGCT
CCCCCGAGGA
TAATGAGCAG
AAACAGCTGT
TTGTGGAACA
GCACAAGCAC
TACCTGGACG
AGATCATCGA
GCAGATCAGC
GAGTTCTCCA
AGAGAGTGAT
CCTGGCCGAC
GCTAATCTGG
ACAAAGTGCT
GTCCGCCTAC
AACAAGCACC
GGGATAAGCC
CATCAGAGAG
CAGGCCGAGA
ATATCATCCA
CCTGTTTACC
CTGACCAATC
TGGGAGCCCC
TGCCGCCTTC
AAGTACTTTG
ACACCACCAT
CGACCGGAAG
AGGTACACCA
GCACCAAAGA
GGTGCTGGAC
GCCACCCTGA
TCCACCAGAG
CATCACCGGC
CTGTACGAGA
CACGGATCGA
CCTGTCTCAG
CTGGGAGGCG
ACAAAAGGCC
GGCGGCCACG
AAAAAGGCCG
GCCAGGCAAA
AAAGAAAAAG
TAAGAATTCC
TAGAGCTCGC
TGATCAGCCT
CGACTGTGCC
TTCTAGTTGC
CAGCCATCTG
TTGTTTGCCC
CTCCCCCGTG
CCTTCCTTGA
CCCTGGAAGG
TGCCACTCCC
ACTGTCCTTT
CCTAATAAAA
TGAGGAAATT
GCATCGCATT
GTCTGAGTAG
GTGTCATTCT
ATTCTGGGGG
GTGGGGTGGG
GCAGGACAGC
AAGGGGGAGG
ATTGGGAAGA
GAATAGCAGG
CATGCTGGGG
AGCGGCCGCA
GGAACCCCTA
GTGATGGAGT
TGGCCACTCC
CTCTCTGCGC
GCTCGCTCGC
TCACTGAGGC
CGGGCGACCA
AAGGTCGCCC
GACGCCCGGG
CTTTGCCCGG
GCGGCCTCAG
TGAGCGAGCG
AGCGCGCAGC
TGCCTGCAGG
GGCGCCTGAT
GCGGTATTTT
CTCCTTACGC
ATCTGTGCGG
TATTTCACAC
CGCATACGTC
AAAGCAACCA
TAGTACGCGC
CCTGTAGCGG
CGCATTAAGC
GCGGCGGGTG
TGGTGGTTAC
GCGCAGCGTG
ACCGCTACAC
TTGCCAGCGC
CCTAGCGCCC
GCTCCTTTCG
CTTTCTTCCC
TTCCTTTCTC
GCCACGTTCG
CCGGCTTTCC
CCGTCAAGCT
CTAAATCGGG
GGCTCCCTTT
AGGGTTCCGA
TTTAGTGCTT
TACGGCACCT
CGACCCCAAA
AAACTTGATT
TGGGTGATGG
TTCACGTAGT
GGGCCATCGC
CCTGATAGAC
GGTTTTTCGC
CCTTTGACGT
TGGAGTCCAC
GTTCTTTAAT
AGTGGACTCT
TGTTCCAAAC
TGGAACAACA
CTCAACCCTA
TCTCGGGCTA
TTCTTTTGAT
TTATAAGGGA
TTTTGCCGAT
TTCGGCCTAT
TGGTTAAAAA
ATGAGCTGAT
TTAACAAAAA
TTTAACGCGA
ATTTTAACAA
AATATTAACG
TTTACAATTT
TATGGTGCAC
TCTCAGTACA
ATCTGCTCTG
ATGCCGCATA
GTTAAGCCAG
CCCCGACACC
CGCCAACACC
CGCTGACGCG
CCCTGACGGG
CTTGTCTGCT
CCCGGCATCC
GCTTACAGAC
AAGCTGTGAC
CGTCTCCGGG
AGCTGCATGT
GTCAGAGGTT
TTCACCGTCA
TCACCGAAAC
GCGCGAGACG
AAAGGGCCTC
GTGATACGCC
TATTTTTATA
GGTTAATGTC
ATGATAATAA
TGGTTTCTTA
GACGTCAGGT
GGCACTTTTC
GGGGAAATGT
GCGCGGAACC
CCTATTTGTT
TATTTTTCTA
AATACATTCA
AATATGTATC
CGCTCATGAG
ACAATAACCC
TGATAAATGC
TTCAATAATA
TTGAAAAAGG
AAGAGTATGA
GTATTCAACA
TTTCCGTGTC
GCCCTTATTC
CCTTTTTTGC
GGCATTTTGC
CTTCCTGTTT
TTGCTCACCC
AGAAACGCTG
GTGAAAGTAA
AAGATGCTGA
AGATCAGTTG
GGTGCACGAG
TGGGTTACAT
CGAACTGGAT
CTCAACAGCG
GTAAGATCCT
TGAGAGTTTT
CGCCCCGAAG
AACGTTTTCC
AATGATGAGC
ACTTTTAAAG
TTCTGCTATG
TGGCGCGGTA
TTATCCCGTA
TTGACGCCGG
GCAAGAGCAA
CTCGGTCGCC
GCATACACTA
TTCTCAGAAT
GACTTGGTTG
AGTACTCACC
AGTCACAGAA
AAGCATCTTA
CGGATGGCAT
GACAGTAAGA
GAATTATGCA
GTGCTGCCAT
AACCATGAGT
GATAACACTG
CGGCCAACTT
ACTTCTGACA
ACGATCGGAG
GACCGAAGGA
GCTAACCGCT
TTTTTGCACA
ACATGGGGGA
TCATGTAACT
CGCCTTGATC
GTTGGGAACC
GGAGCTGAAT
GAAGCCATAC
CAAACGACGA
GCGTGACACC
ACGATGCCTG
TAGCAATGGC
AACAACGTTG
CGCAAACTAT
TAACTGGCGA
ACTACTTACT
CTAGCTTCCC
GGCAACAATT
AATAGACTGG
ATGGAGGCGG
ATAAAGTTGC
AGGACCACTT
CTGCGCTCGG
CCCTTCCGGC
TGGCTGGTTT
ATTGCTGATA
AATCTGGAGC
CGGTGAGCGT
GGAAGCCGCG
GTATCATTGC
AGCACTGGGG
CCAGATGGTA
AGCCCTCCCG
TATCGTAGTT
ATCTACACGA
CGGGGAGTCA
GGCAACTATG
GATGAACGAA
ATAGACAGAT
CGCTGAGATA
GGTGCCTCAC
TGATTAAGCA
TTGGTAACTG
TCAGACCAAG
TTTACTCATA
TATACTTTAG
ATTGATTTAA
AACTTCATTT
TTAATTTAAA
AGGATCTAGG
TGAAGATCCT
TTTTGATAAT
CTCATGACCA
AAATCCCTTA
ACGTGAGTTT
TCGTTCCACT
GAGCGTCAGA
CCCCGTAGAA
AAGATCAAAG
GATCTTCTTG
AGATCCTTTT
TTTCTGCGCG
TAATCTGCTG
CTTGCAAACA
AAAAAACCAC
CGCTACCAGC
GGTGGTTTGT
TTGCCGGATC
AAGAGCTACC
AACTCTTTTT
CCGAAGGTAA
CTGGCTTCAG
CAGAGCGCAG
ATACCAAATA
CTGTCCTTCT
AGTGTAGCCG
TAGTTAGGCC
ACCACTTCAA
GAACTCTGTA
GCACCGCCTA
CATACCTCGC
TCTGCTAATC
CTGTTACCAG
TGGCTGCTGC
CAGTGGCGAT
AAGTCGTGTC
TTACCGGGTT
GGACTCAAGA
CGATAGTTAC
CGGATAAGGC
GCAGCGGTCG
GGCTGAACGG
GGGGTTCGTG
CACACAGCCC
AGCTTGGAGC
GAACGACCTA
CACCGAACTG
AGATACCTAC
AGCGTGAGCT
ATGAGAAAGC
GCCACGCTTC
CCGAAGGGAG
AAAGGCGGAC
AGGTATCCGG
TAAGCGGCAG
GGTCGGAACA
GGAGAGCGCA
CGAGGGAGCT
TCCAGGGGGA
AACGCCTGGT
ATCTTTATAG
TCCTGTCGGG
TTTCGCCACC
TCTGACTTGA
GCGTCGATTT
TTGTGATGCT
CGTCAGGGGG
GCGGAGCCTA
TGGAAAAACG
CCAGCAACGC
GGCCTTTTTA
CGGTTCCTGG
CCTTTTGCTG
GCCTTTTGCT
CACATGT