名称: | pCS2-nCas9n |
---|---|
别称: | Plasmid#47929 |
基因长: | |
质粒宿主: | 哺乳细胞,鱼类 |
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: | CMV |
复制子: | pUC |
拷贝数: | |
感受态: | DH5a |
温度: | 37度 |
背景载体: | pCS2+ |
正向引物: | CMV-F: |
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。
CGCCATTCTG
CCTGGGGACG
TCGGAGCAAG
CTTGATTTAG
GTGACACTAT
AGAATACAAG
CTACTTGTTC
TTTTTGCAGG
ATCCGATCCA
ATGTCGACGC
CACCATGGCT
TCTCCACCTA
AGAAGAAGAG
AAAGGTGGGA
AGCATGGATA
AGAAGTATAG
CATCGGCCTG
GATATTGGAA
CTAACTCCGT
GGGTTGGGCA
GTGATTACAG
ACGACTACAA
GGTCCCTAGC
AAGAAATTTA
AGGTGCTGGG
TAACACCGAC
AGGCACAGCA
TCAAGAAAAA
TCTGATTGGA
GCCCTGCTGT
TCGGTTCTGG
AGAGACTGCC
GAAGCAACAC
GCCTGAAAAG
AACAGCAAGA
AGGCGCTATA
CCAGAAGGAA
GAATAGAATC
TGTTACCTGC
AGGAGATTTT
CTCTAACGAA
ATGGCTAAGG
TGGACGATTC
ATTCTTTCAT
AGGCTGGAGG
AAAGTTTCCT
GGTCGAGGAA
GATAAGAAAC
ACGAGCGCCA
TCCTATCTTT
GGAAACATTG
TGGACGAGGT
CGCCTATCAC
GAAAAATACC
CAACCATCTA
TCATCTGCGC
AAGAAACTGG
CTGACTCTAC
TGATAAAGCC
GACCTGAGAC
TGATCTATCT
GGCTCTGGCC
CACATGATTA
AGTTCAGGGG
TCATTTTCTG
ATCGAGGGCG
ATCTGAACCC
CGACAATTCC
GATGTGGACA
AGCTGTTCAT
CCAGCTGGTC
CAGATTTACA
ATCAGCTGTT
TGAGGAAAAC
CCTATTAATG
CTTCCAGAGT
GGACGCAAAA
GCTATCCTGT
CAGCCAGGCT
GTCCAAGTCA
CGCAGACTGG
AGAACCTGAT
TGCACAGCTG
CCCGGAGAAA
AGAGGAACGG
TCTGTTTGGA
AATCTGATCG
CTCTGAGTCT
GGGCCTGACT
CCTAACTTCA
AAAGCAATTT
TGATCTGGCT
GAGGACGCCA
AACTGCAGCT
GTCAAAGGAC
ACATATGACG
ATGACCTGGA
TAACCTGCTG
GCACAGATCG
GAGATCAGTA
CGCTGACCTG
TTCCTGGCTG
CCAAAAATCT
GTCCGACGCA
ATCCTGCTGT
CAGATATTCT
GAGAGTGAAC
AGCGAGATTA
CAAAAGCTCC
TCTGAGTGCC
AGCATGATCA
AGAGATATGA
CGAGCACCAT
CAGGATCTGA
CCCTGCTGAA
GGCTCTGGTC
AGGCAGCAGC
TGCCAGAGAA
GTACAAGGAA
ATTTTCTTTG
ATCAGTCCAA
GAACGGCTAC
GCCGGTTATA
TCGACGGAGG
CGCATCACAG
GAGGAATTCT
ACAAGTTTAT
CAAACCTATT
CTGGAGAAGA
TGGACGGAAC
TGAGGAACTG
CTGGTGAAAC
TGAATAGAGA
GGACCTGCTG
AGGAAGCAGC
GCACATTTGA
TAACGGTTCC
ATCCCACACC
AGATTCATCT
GGGAGAGCTG
CACGCTATCC
TGAGGCGCCA
GGAAGACTTC
TACCCCTTTC
TGAAAGATAA
CCGCGAGAAG
ATCGAAAAAA
TTCTGACCTT
CAGAATCCCT
TACTATGTGG
GTCCACTGGC
TCGCGGAAAC
AGCAGATTTG
CCTGGATGAC
TCGCAAATCC
GAGGAAACCA
TTACTCCTTG
GAACTTCGAG
GAAGTGGTCG
ATAAGGGCGC
CTCTGCACAG
TCCTTCATCG
AGAGAATGAC
TAATTTTGAC
AAAAACCTGC
CCAATGAGAA
AGTGCTGCCT
AAGCACTCCC
TGCTGTACGA
GTATTTCACT
GTCTATAACG
AACTGACAAA
GGTGAAATAC
GTCACCGAGG
GCATGAGAAA
GCCAGCCTTC
CTGTCAGGAG
AGCAGAAGAA
AGCAATCGTG
GATCTGCTGT
TTAAAACCAA
CAGGAAAGTG
ACTGTCAAGC
AGCTGAAGGA
GGACTACTTC
AAGAAAATTG
AATGCTTCGA
TTCCGTGGAG
ATCAGCGGAG
TCGAAGACAG
ATTTAACGCA
AGCCTGGGCG
CTTACCACGA
TCTGCTGAAG
ATCATTAAGG
ATAAAGACTT
CCTGGACAAC
GAGGAAAATG
AGGATATCCT
GGAAGACATT
GTGCTGACAC
TGACCCTGTT
TGAGGACAGA
GGAATGATCG
AGGAAAGACT
GAAAACCTAT
GCTCATCTGT
TCGATGACAA
GGTGATGAAA
CAGCTGAAGA
GAAGGCGCTA
CACTGGCTGG
GGTAGACTGA
GCAGGAAGCT
GATCAACGGC
ATTAGGGATA
AACAGTCAGG
AAAGACAATC
CTGGACTTTC
TGAAAAGTGA
TGGCTTCGCC
AACCGCAATT
TTATGCAGCT
GATTCACGAT
GACAGTCTGA
CCTTCAAAGA
GGACATCCAG
AAGGCTCAGG
TGTCTGGACA
GGGCCACTCC
CTGCATGAGC
AGATTGCAAA
CCTGGCTGGA
AGCCCAGCCA
TCAAGAAAGG
CATTCTGCAG
ACAGTGAAAA
TCGTCGATGA
GCTGGTGAAA
GTCATGGGCC
ATAAGCCCGA
AAACATCGTG
ATTGAGATGG
CTCGCGAAAA
TCAGACAACC
CAGAAGGGTC
AGAAGAACAG
TAGAGAGAGG
ATGAAAAGAA
TCGAGGAAGG
CATTAAGGAG
CTGGGTAGCC
AGATCCTGAA
AGAGCACCCA
GTGGAAAACA
CACAGCTGCA
GAATGAGAAG
CTGTATCTGT
ACTATCTGCA
GAATGGAAGA
GATATGTACG
TGGACCAGGA
GCTGGATATT
AACAGGCTGT
CTGATTACGA
CGTGGATCAT
ATCGTCCCCC
AGAGTTTCAT
CAAAGATGAC
AGCATTGACA
ACAAGGTGCT
GACCAGGTCC
GACAAAAACA
GAGGAAAATC
AGATAATGTC
CCTAGTGAGG
AAGTGGTCAA
GAAAATGAAG
AACTACTGGA
GACAGCTGCT
GAATGCCAAA
CTGATCACTC
AGAGGAAGTT
TGATAACCTG
ACAAAAGCAG
AGCGCGGTGG
ACTGTCAGAA
CTGGACAAAG
CTGGATTCAT
CAAGAGGCAG
CTGGTGGAAA
CACGCCAGAT
CACTAAACAC
GTCGCACAGA
TTCTGGATAG
TCGCATGAAC
ACAAAGTACG
ATGAGAATGA
CAAACTGATC
AGAGAAGTGA
AGGTCATTAC
CCTGAAGAGT
AAACTGGTCA
GCGACTTTAG
GAAAGATTTC
CAGTTTTATA
AGGTCCGCGA
GATTAACAAT
TATCACCATG
CCCATGACGC
ATACCTGAAC
GCCGTGGTCG
GTACCGCACT
GATCAAGAAA
TACCCAAAAC
TGGAGAGCGA
ATTCGTGTAC
GGAGACTATA
AGGTGTACGA
TGTCAGAAAA
ATGATCGCCA
AGTCCGAGCA
GGAAATTGGA
AAAGCTACTG
CCAAGTATTT
CTTTTACTCA
AACATCATGA
ATTTCTTTAA
GACAGAGATC
ACCCTGGCCA
ATGGAGAAAT
CCGCAAAAGG
CCCCTGATTG
AGACAAACGG
AGAGACAGGC
GAAATCGTGT
GGGACAAAGG
CAGAGATTTT
GCAACCGTGA
GGAAGGTCCT
GAGCATGCCT
CAAGTGAATA
TCGTCAAGAA
AACTGAGGTG
CAGACAGGCG
GTTTCTCAAA
AGAAAGTATT
CTGCCAAAAC
GCAACTCTGA
TAAGCTGATC
GCTAGAAAGA
AAGACTGGGA
CCCTAAGAAG
TATGGAGGCT
TTGACTCTCC
CACTGTGGCA
TACTCCGTCC
TGGTGGTCGC
TAAGGTGGAG
AAGGGCAAAA
GCAAGAAACT
GAAATCTGTC
AAGGAGCTGC
TGGGTATCAC
AATTATGGAG
AGAAGCTCTT
TCGAGAAGAA
CCCAATCGAT
TTTCTGGAGG
CCAAAGGTTA
TAAGGAAGTG
AAGAAAGACC
TGATCATTAA
ACTGCCCAAG
TACAGTCTGT
TTGAGCTGGA
AAACGGCAGG
AAACGCATGC
TGGCAAGCGC
TGGAGAGCTG
CAGAAAGGCA
ATGAACTGGC
CCTGCCTTCT
AAGTACGTGA
ACTTCCTGTA
TCTGGCAAGC
CACTACGAGA
AGCTGAAAGG
ATCTCCAGAG
GATAACGAAC
AGAAACAGCT
GTTTGTGGAG
CAGCACAAGC
ATTATCTGGA
CGAGATCATT
GAACAGATTA
GCGAGTTCTC
TAAAAGAGTG
ATCCTGGCCG
ACGCAAATCT
GGATAAGGTC
CTGTCTGCTT
ACAACAAACA
CAGAGATAAG
CCCATCAGGG
AGCAGGCCGA
AAATATCATT
CATCTGTTCA
CTCTGACAAA
CCTGGGCGCA
CCTGCAGCTT
TCAAGTACTT
CGACACTACA
ATCGATAGAA
AGAGGTACAC
CTCCACTAAG
GAGGTGCTGG
ACGCTACACT
GATCCATCAG
AGTATTACCG
GCCTGTACGA
AACAAGGATT
GACCTGTCTC
AGCTGGGTGG
CGACTCACCG
GTGAGATCTC
CTAAGAAGAA
GAGAAAGGTG
TGACCGCGGA
ATCTAGAACT
ATAGTGAGTC
GTATTACGTA
GATCCAGACA
TGATAAGATA
CATTGATGAG
TTTGGACAAA
CCACAACTAG
AATGCAGTGA
AAAAAATGCT
TTATTTGTGA
AATTTGTGAT
GCTATTGCTT
TATTTGTAAC
CATTATAAGC
TGCAATAAAC
AAGTTAACAA
CAACAATTGC
ATTCATTTTA
TGTTTCAGGT
TCAGGGGGAG
GTGTGGGAGG
TTTTTTAATT
CGCGGCCGCG
GCGCCAATGC
ATTGGGCCCG
GTACCCAGCT
TTTGTTCCCT
TTAGTGAGGG
TTAATTGCGC
GCTTGGCGTA
ATCATGGTCA
TAGCTGTTTC
CTGTGTGAAA
TTGTTATCCG
CTCACAATTC
CACACAACAT
ACGAGCCGGA
AGCATAAAGT
GTAAAGCCTG
GGGTGCCTAA
TGAGTGAGCT
AACTCACATT
AATTGCGTTG
CGCTCACTGC
CCGCTTTCCA
GTCGGGAAAC
CTGTCGTGCC
AGCTGCATTA
ATGAATCGGC
CAACGCGCGG
GGAGAGGCGG
TTTGCGTATT
GGGCGCTCTT
CCGCTTCCTC
GCTCACTGAC
TCGCTGCGCT
CGGTCGTTCG
GCTGCGGCGA
GCGGTATCAG
CTCACTCAAA
GGCGGTAATA
CGGTTATCCA
CAGAATCAGG
GGATAACGCA
GGAAAGAACA
TGTGAGCAAA
AGGCCAGCAA
AAGGCCAGGA
ACCGTAAAAA
GGCCGCGTTG
CTGGCGTTTT
TCCATAGGCT
CCGCCCCCCT
GACGAGCATC
ACAAAAATCG
ACGCTCAAGT
CAGAGGTGGC
GAAACCCGAC
AGGACTATAA
AGATACCAGG
CGTTTCCCCC
TGGAAGCTCC
CTCGTGCGCT
CTCCTGTTCC
GACCCTGCCG
CTTACCGGAT
ACCTGTCCGC
CTTTCTCCCT
TCGGGAAGCG
TGGCGCTTTC
TCATAGCTCA
CGCTGTAGGT
ATCTCAGTTC
GGTGTAGGTC
GTTCGCTCCA
AGCTGGGCTG
TGTGCACGAA
CCCCCCGTTC
AGCCCGACCG
CTGCGCCTTA
TCCGGTAACT
ATCGTCTTGA
GTCCAACCCG
GTAAGACACG
ACTTATCGCC
ACTGGCAGCA
GCCACTGGTA
ACAGGATTAG
CAGAGCGAGG
TATGTAGGCG
GTGCTACAGA
GTTCTTGAAG
TGGTGGCCTA
ACTACGGCTA
CACTAGAAGG
ACAGTATTTG
GTATCTGCGC
TCTGCTGAAG
CCAGTTACCT
TCGGAAAAAG
AGTTGGTAGC
TCTTGATCCG
GCAAACAAAC
CACCGCTGGT
AGCGGTGGTT
TTTTTGTTTG
CAAGCAGCAG
ATTACGCGCA
GAAAAAAAGG
ATCTCAAGAA
GATCCTTTGA
TCTTTTCTAC
GGGGTCTGAC
GCTCAGTGGA
ACGAAAACTC
ACGTTAAGGG
ATTTTGGTCA
TGAGATTATC
AAAAAGGATC
TTCACCTAGA
TCCTTTTAAA
TTAAAAATGA
AGTTTTAAAT
CAATCTAAAG
TATATATGAG
TAAACTTGGT
CTGACAGTTA
CCAATGCTTA
ATCAGTGAGG
CACCTATCTC
AGCGATCTGT
CTATTTCGTT
CATCCATAGT
TGCCTGACTC
CCCGTCGTGT
AGATAACTAC
GATACGGGAG
GGCTTACCAT
CTGGCCCCAG
TGCTGCAATG
ATACCGCGAG
ACCCACGCTC
ACCGGCTCCA
GATTTATCAG
CAATAAACCA
GCCAGCCGGA
AGGGCCGAGC
GCAGAAGTGG
TCCTGCAACT
TTATCCGCCT
CCATCCAGTC
TATTAATTGT
TGCCGGGAAG
CTAGAGTAAG
TAGTTCGCCA
GTTAATAGTT
TGCGCAACGT
TGTTGCCATT
GCTACAGGCA
TCGTGGTGTC
ACGCTCGTCG
TTTGGTATGG
CTTCATTCAG
CTCCGGTTCC
CAACGATCAA
GGCGAGTTAC
ATGATCCCCC
ATGTTGTGCA
AAAAAGCGGT
TAGCTCCTTC
GGTCCTCCGA
TCGTTGTCAG
AAGTAAGTTG
GCCGCAGTGT
TATCACTCAT
GGTTATGGCA
GCACTGCATA
ATTCTCTTAC
TGTCATGCCA
TCCGTAAGAT
GCTTTTCTGT
GACTGGTGAG
TACTCAACCA
AGTCATTCTG
AGAATAGTGT
ATGCGGCGAC
CGAGTTGCTC
TTGCCCGGCG
TCAATACGGG
ATAATACCGC
GCCACATAGC
AGAACTTTAA
AAGTGCTCAT
CATTGGAAAA
CGTTCTTCGG
GGCGAAAACT
CTCAAGGATC
TTACCGCTGT
TGAGATCCAG
TTCGATGTAA
CCCACTCGTG
CACCCAACTG
ATCTTCAGCA
TCTTTTACTT
TCACCAGCGT
TTCTGGGTGA
GCAAAAACAG
GAAGGCAAAA
TGCCGCAAAA
AAGGGAATAA
GGGCGACACG
GAAATGTTGA
ATACTCATAC
TCTTCCTTTT
TCAATATTAT
TGAAGCATTT
ATCAGGGTTA
TTGTCTCATG
AGCGGATACA
TATTTGAATG
TATTTAGAAA
AATAAACAAA
TAGGGGTTCC
GCGCACATTT
CCCCGAAAAG
TGCCACCTAA
ATTGTAAGCG
TTAATATTTT
GTTAAAATTC
GCGTTAAATT
TTTGTTAAAT
CAGCTCATTT
TTTAACCAAT
AGGCCGAAAT
CGGCAAAATC
CCTTATAAAT
CAAAAGAATA
GACCGAGATA
GGGTTGAGTG
TTGTTCCAGT
TTGGAACAAG
AGTCCACTAT
TAAAGAACGT
GGACTCCAAC
GTCAAAGGGC
GAAAAACCGT
CTATCAGGGC
GATGGCCCAC
TACGTGAACC
ATCACCCTAA
TCAAGTTTTT
TGGGGTCGAG
GTGCCGTAAA
GCACTAAATC
GGAACCCTAA
AGGGAGCCCC
CGATTTAGAG
CTTGACGGGG
AAAGCCGGCG
AACGTGGCGA
GAAAGGAAGG
GAAGAAAGCG
AAAGGAGCGG
GCGCTAGGGC
GCTGGCAAGT
GTAGCGGTCA
CGCTGCGCGT
AACCACCACA
CCCGCCGCGC
TTAATGCGCC
GCTACAGGGC
GCGTCCCATT
CGCCATTCAG
GCTGCGCAAC
TGTTGGGAAG
GGCGATCGGT
GCGGGCCTCT
TCGCTATTAC
GCCAGTCGAC
CATAGCCAAT
TCAATATGGC
GTATATGGAC
TCATGCCAAT
TCAATATGGT
GGATCTGGAC
CTGTGCCAAT
TCAATATGGC
GTATATGGAC
TCGTGCCAAT
TCAATATGGT
GGATCTGGAC
CCCAGCCAAT
TCAATATGGC
GGACTTGGCA
CCATGCCAAT
TCAATATGGC
GGACTTGGCA
CTGTGCCAAC
TGGGGAGGGG
TCTACTTGGC
ACGGTGCCAA
GTTTGAGGAG
GGGTCTTGGC
CCTGTGCCAA
GTCCGCCATA
TTGAATTGGC
ATGGTGCCAA
TAATGGCGGC
CATATTGGCT
ATATGCCAGG
ATCAATATAT
AGGCAATATC
CAATATGGCC
CTATGCCAAT
ATGGCTATTG
GCCAGGTTCA
ATACTATGTA
TTGGCCCTAT
GCCATATAGT
ATTCCATATA
TGGGTTTTCC
TATTGACGTA
GATAGCCCCT
CCCAATGGGC
GGTCCCATAT
ACCATATATG
GGGCTTCCTA
ATACCGCCCA
TAGCCACTCC
CCCATTGACG
TCAATGGTCT
CTATATATGG
TCTTTCCTAT
TGACGTCATA
TGGGCGGTCC
TATTGACGTA
TATGGCGCCT
CCCCCATTGA
CGTCAATTAC
GGTAAATGGC
CCGCCTGGCT
CAATGCCCAT
TGACGTCAAT
AGGACCACCC
ACCATTGACG
TCAATGGGAT
GGCTCATTGC
CCATTCATAT
CCGTTCTCAC
GCCCCCTATT
GACGTCAATG
ACGGTAAATG
GCCCACTTGG
CAGTACATCA
ATATCTATTA
ATAGTAACTT
GGCAAGTACA
TTACTATTGG
AAGGACGCCA
GGGTACATTG
GCAGTACTCC
CATTGACGTC
AATGGCGGTA
AATGGCCCGC
GATGGCTGCC
AAGTACATCC
CCATTGACGT
CAATGGGGAG
GGGCAATGAC
GCAAATGGGC
GTTCCATTGA
CGTAAATGGG
CGGTAGGCGT
GCCTAATGGG
AGGTCTATAT
AAGCAATGCT
CGTTTAGGGA
AC