名称: | pEJS654 All-in-One AAV-sgRNA-hNmeCas9 |
---|---|
别称: | Plasmid#112139 |
基因长: | |
质粒宿主: | 哺乳细胞,腺相关病毒 |
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。
CCTGCAGGCA
GCTGCGCGCT
CGCTCGCTCA
CTGAGGCCGC
CCGGGCGTCG
GGCGACCTTT
GGTCGCCCGG
CCTCAGTGAG
CGAGCGAGCG
CGCAGAGAGG
GAGTGGCCAA
CTCCATCACT
AGGGGTTCCT
GCGGCCTCTA
GAGTTTAAAC
AAAAAAATAA
ACGATGCCCC
TTAAAGCAGA
AGCTTTAAGG
GGCAGAGCGT
TGCGGCACAT
CTTTTCAGAC
GGCCTTATTG
TAGCAACGGT
TCTCATTTCG
TTTCCGAAAT
GAGAAAGGGA
GCTACAACTG
AAGAGCTGGA
ATGCTCTTCC
GGTGTTTCGT
CCTTTCCACA
AGATATATAA
AGCCAAGAAA
TCGAAATACT
TTCAAGTTAC
GGTAAGCATA
TGATAGTCCA
TTTTAAAACA
TAATTTTAAA
ACTGCAAACT
ACCCAAGAAA
TTATTACTTT
CTACGTCACG
TATTTTGTAC
TAATATCTTT
GTGTTTACAG
TCAAATTAAT
TCTAATTATC
TCTCTAACAG
CCTTGTATCG
TATATGCAAA
TATGAAGGAA
TCATGGGAAA
TAGGCCCTCT
TCCTGCCCGA
CCTTGACGTC
GACTCTAGAA
TGGAGGCGGT
ACTATGTAGA
TGAGAATTCA
GGAGCAAACT
GGGAAAAGCA
ACTGCTTCCA
AATATTTGTG
ATTTTTACAG
TGTAGTTTTG
GAAAAACTCT
TAGCCTACCA
ATTCTTCTAA
GTGTTTTAAA
ATGTGGGAGC
CAGTACACAT
GAAGTTATAG
AGTGTTTTAA
TGAGGCTTAA
ATATTTACCG
TAACTATGAA
ATGCTACGCA
TATCATGCTG
TTCAGGCTCC
GTGGCCACGC
AACTCATACT
TAAGCAGACA
GTGGTTCAAA
GTTTTTTTCT
TCCATTTCAG
GTGTCGTGAA
CACCGCCACC
ATGGTGCCTA
AGAAGAAGAG
AAAGGTGGAA
GATAAACGCC
CAGCAGCTAC
AAAGAAGGCA
GGTCAAGCCA
AGAAAAAGAA
AGCAGCATTC
AAGCCAAACT
CAATCAATTA
CATCCTGGGA
CTGGACATCG
GCATCGCATC
CGTCGGGTGG
GCTATGGTCG
AAATCGACGA
GGAGGAGAAC
CCCATCCGCC
TGATCGATCT
GGGCGTGCGC
GTGTTTGAGA
GGGCAGAGGT
GCCTAAGACC
GGCGACAGCC
TGGCCATGGC
ACGGAGACTG
GCACGCTCCG
TGAGGCGCCT
GACCCGGAGA
AGGGCCCACA
GACTGCTGAG
GACACGCCGG
CTGCTGAAGA
GGGAGGGCGT
GCTGCAGGCC
GCCAACTTCG
ATGAGAATGG
CCTGATCAAG
TCCCTGCCCA
ATACCCCTTG
GCAGCTGAGG
GCAGCCGCCC
TGGACCGCAA
GCTGACACCT
CTGGAGTGGT
CCGCCGTGCT
GCTGCACCTG
ATCAAGCACC
GGGGCTACCT
GTCTCAGAGA
AAGAACGAGG
GCGAGACAGC
CGATAAGGAG
CTGGGCGCCC
TGCTGAAGGG
AGTGGCAGGA
AATGCACACG
CCCTGCAGAC
CGGCGACTTT
CGCACACCAG
CCGAGCTGGC
CCTGAACAAG
TTCGAGAAGG
AGAGCGGCCA
CATCCGCAAT
CAGCGGTCTG
ACTATAGCCA
CACCTTCTCC
CGGAAGGATC
TGCAGGCCGA
GCTGATCCTG
CTGTTTGAGA
AGCAGAAGGA
GTTCGGCAAC
CCACACGTGT
CTGGCGGCCT
GAAGGAGGGC
ATCGAGACAC
TGCTGATGAC
ACAGCGGCCC
GCCCTGAGCG
GCGACGCAGT
GCAGAAGATG
CTGGGACACT
GCACCTTTGA
GCCAGCCGAG
CCCAAGGCCG
CCAAGAATAC
CTACACAGCC
GAGCGGTTCA
TCTGGCTGAC
AAAGCTGAAC
AATCTGAGGA
TCCTGGAGCA
GGGAAGCGAG
CGCCCACTGA
CCGACACAGA
GAGGGCCACC
CTGATGGATG
AGCCCTACCG
CAAGTCCAAG
CTGACATATG
CACAGGCAAG
GAAGCTGCTG
GGCCTGGAGG
ACACCGCCTT
CTTTAAGGGC
CTGAGATACG
GCAAGGATAA
CGCCGAGGCC
TCTACACTGA
TGGAGATGAA
GGCCTATCAC
GCCATCAGCA
GGGCCCTGGA
GAAGGAGGGC
CTGAAGGACA
AGAAGTCCCC
ACTGAATCTG
TCTCCCGAGC
TGCAGGATGA
GATCGGCACC
GCCTTTAGCC
TGTTCAAGAC
CGACGAGGAT
ATCACAGGCA
GACTGAAGGA
CAGGATCCAG
CCAGAGATCC
TGGAGGCCCT
GCTGAAGCAC
ATCAGCTTTG
ATAAGTTCGT
GCAGATCAGC
CTGAAGGCCC
TGCGGAGGAT
CGTGCCACTG
ATGGAGCAGG
GCAAGAGGTA
CGACGAGGCC
TGCGCCGAAA
TCTACGGCGA
TCACTATGGC
AAGAAGAACA
CAGAGGAGAA
AATCTACCTG
CCCCCTATCC
CCGCCGATGA
GATCAGGAAC
CCTGTGGTGC
TGCGCGCCCT
GTCTCAGGCA
AGAAAAGTGA
TCAACGGAGT
GGTGCGCCGG
TACGGCAGCC
CCGCCAGAAT
CCACATCGAG
ACAGCCAGGG
AAGTGGGCAA
GTCCTTTAAG
GACAGAAAGG
AGATCGAGAA
GAGGCAGGAG
GAGAACAGAA
AGGATAGGGA
GAAGGCCGCC
GCCAAGTTCA
GAGAGTACTT
TCCTAATTTC
GTGGGCGAGC
CAAAGTCCAA
GGATATCCTG
AAGCTGAGGC
TGTACGAGCA
GCAGCACGGC
AAGTGTCTGT
ATTCTGGCAA
GGAGATCAAC
CTGGGCCGCC
TGAATGAGAA
GGGCTATGTG
GAGATCGACC
ACGCCCTGCC
TTTTTCTCGG
ACCTGGGACG
ATAGCTTCAA
CAATAAGGTG
CTGGTGCTGG
GCTCTGAGAA
CCAGAATAAG
GGCAACCAGA
CACCCTACGA
GTATTTCAAC
GGCAAGGACA
ATAGCCGCGA
GTGGCAGGAG
TTTAAGGCAA
GGGTGGAGAC
AAGCAGGTTC
CCTCGGTCCA
AGAAGCAGAG
AATCCTGCTG
CAGAAGTTTG
ACGAGGATGG
CTTCAAGGAG
AGGAACCTGA
ATGACACCCG
CTACGTGAAT
CGGTTTCTGT
GCCAGTTCGT
GGCCGATAGA
ATGAGGCTGA
CCGGCAAGGG
CAAGAAGAGA
GTGTTTGCCT
CCAACGGCCA
GATCACAAAT
CTGCTGAGGG
GCTTCTGGGG
CCTGAGAAAG
GTGAGGGCAG
AGAACGACAG
GCACCACGCA
CTGGATGCAG
TGGTGGTGGC
ATGTTCTACC
GTGGCCATGC
AGCAGAAGAT
CACACGCTTT
GTGCGGTATA
AGGAGATGAA
TGCCTTCGAC
GGCAAGACCA
TCGATAAGGA
GACAGGCGAG
GTGCTGCACC
AGAAGACACA
CTTTCCTCAG
CCATGGGAGT
TCTTTGCCCA
GGAAGTGATG
ATCCGGGTGT
TTGGCAAGCC
TGACGGCAAG
CCAGAGTTCG
AGGAGGCCGA
TACCCTGGAG
AAGCTGAGAA
CACTGCTGGC
AGAGAAGCTG
AGCTCCAGGC
CCGAGGCAGT
GCACGAGTAC
GTGACCCCAC
TGTTCGTGTC
TAGAGCCCCC
AACAGGAAGA
TGAGCGGCCA
GGGCCACATG
GAGACAGTGA
AGTCCGCCAA
GAGACTGGAC
GAGGGCGTGT
CTGTGCTGAG
GGTGCCTCTG
ACACAGCTGA
AGCTGAAGGA
TCTGGAGAAG
ATGGTGAATC
GCGAGCGGGA
GCCAAAGCTG
TATGAGGCCC
TGAAGGCAAG
GCTGGAGGCA
CACAAGGACG
ATCCTGCCAA
GGCCTTTGCC
GAGCCATTCT
ACAAGTATGA
TAAGGCCGGC
AACAGAACCC
AGCAGGTGAA
GGCCGTGAGG
GTGGAGCAGG
TGCAGAAGAC
AGGCGTGTGG
GTGCGCAACC
ACAATGGCAT
CGCCGACAAT
GCTACCATGG
TGCGGGTGGA
CGTGTTTGAG
AAGGGCGATA
AGTACTATCT
GGTGCCCATC
TACAGCTGGC
AGGTGGCCAA
GGGCATCCTG
CCTGATAGAG
CCGTGGTGCA
GGGCAAGGAC
GAGGAGGATT
GGCAGCTGAT
CGACGATTCC
TTCAACTTTA
AGTTCTCTCT
GCACCCCAAT
GACCTGGTGG
AAGTGATCAC
CAAGAAGGCC
AGGATGTTTG
GCTACTTCGC
CTCCTGCCAC
CGCGGCACAG
GCAACATCAA
TATCCGGATC
CACGACCTGG
ATCACAAGAT
CGGCAAGAAC
GGCATCCTGG
AGGGCATCGG
CGTGAAGACA
GCCCTGAGCT
TCCAGAAGTA
TCAGATCGAC
GAGCTGGGCA
AGGAGATCAG
ACCTTGTAGG
CTGAAGAAGC
GCCCACCCGT
GCGGGAGGAT
AAGCGGCCCG
CAGCAACCAA
GAAGGCAGGA
CAGGCCAAGA
AGAAGAAGTA
CCCCTATGAC
GTGCCTGACT
ACGCCGGGTA
TCCCTACGAC
GTGCCTGATT
ACGCCGGGTC
CTATCCCTAC
GACGTGCCAG
ATTACGCTGC
AGCTCCAGCA
GCGAAGAAAA
AGAAGCTGGA
TTAAGATCTT
TTTCCCTCTG
CCAAAAATTA
TGGGGACATC
ATGAAGCCCC
TTGAGCATCT
GACTTCTGGC
TAATAAAGGA
AATTTATTTT
CATTGCAATA
GTGTGTTGGA
ATTTTTTGTG
TCTCTCACTC
GGCGGCCGCA
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
CTTAGCGCCC
GCTCCTTTCG
CTTTCTTCCC
TTCCTTTCTC
GCCACGTTCG
CCGGCTTTCC
CCGTCAAGCT
CTAAATCGGG
GGCTCCCTTT
AGGGTTCCGA
TTTAGTGCTT
TACGGCACCT
CGACCCCAAA
AAACTTGATT
TGGGTGATGG
TTCACGTAGT
GGGCCATCGC
CCTGATAGAC
GGTTTTTCGC
CCTTTGACGT
TGGAGTCCAC
GTTCTTTAAT
AGTGGACTCT
TGTTCCAAAC
TGGAACAACA
CTCAACTCTA
TCTCGGGCTA
TTCTTTTGAT
TTATAAGGGA
TTTTGCCGAT
TTCGGTCTAT
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
CTGTTCTTCT
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