名称: | plRL2 |
---|---|
别称: | Plasmid#163631 |
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: | 空载体 |
原核抗性: | Kan |
筛选标记: | |
荧光标记: | |
启动子: | Tet-On |
复制子: | pUC |
拷贝数: | 低 |
感受态: | DH5a |
温度: | 37度 |
背景载体: | |
正向引物: | Forward(left ITR) |
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。
AATAAACGAA
AGGCTCAGTC
GAAAGACTGG
GCCTTTCGTT
TATCTGTTGT
TTGAAAAAAA
AAAGCGCCGC
AACTGCGGCG
CTTTTTTTTT
TGAATTCTCT
GACCAGGGAA
AATAGCCCTC
TGACCTGGGG
ATTTGCGATC
CCTATCAGTG
ATAGATATAA
TCTGGGAGGA
GACGATTAAT
GCGTCTCGGT
TTTTGTACTC
GAAAGAAGCT
ACAAAGATAA
GGCTTCATGC
CGAAATCAAC
ACCCTGTCAT
TTTATGGCAG
GGTGTTTTTT
TTTTGTCGAC
TTGGGGACCC
TAGAGGTCCC
CTTTTTTATT
TTTTTGGATG
AAGAGCATTA
TCAATGGCTC
TTCACAGGAA
GCTTAAAAAA
AAAAGCCCCG
CGATTGCGGG
GCTTTTTTTT
TTTAATACGG
TTATCCACAG
AATCAGGGGA
TAACGCAGGA
AAGAACATGT
GAGCAAAAGG
CCAGCAAAAG
GCCAGGAACC
GTAAAAAGGC
CGCGTTGCTG
GCGTTTTTCC
ATAGGCTCCG
CCCCCCTGAC
GAGCATCACA
AAAATCGACG
CTCAAGTCAG
AGGTGGCGAA
ACCCGACAGG
ACTATAAAGA
TACCAGGCGT
TTCCCCCTGG
AAGCTCCCTC
GTGCGCTCTC
CTGTTCCGAC
CCTGCCGCTT
ACCGGATACC
TGTCCGCCTT
TCTCCCCTCG
GGAAGCGTGG
CGCTTTCTCA
TAGCTCACGC
TGTAGGTATC
TCAGTTCGGT
GTAGGTCGTT
CGCTCCAAGC
TGGGCTGTGT
GCACGAACCC
CCCGTTCAGC
CCGACCGCTG
CGCCTTATCC
GGTAACTATC
GTCTTGAGTC
CAACCCGGTA
AGACACGACT
TATCGCCACT
GGCAGCAGCC
ACTGGTAACA
GGATTAGCAG
AGCGAGGTAT
GTAGGCGGTG
CTACAGAGTT
CTTGAAGTGG
TGGCCTAACT
ACGGCTACAC
TAGAAGGACA
GTATTTGGTA
TCTGCGCTCT
GCTGAAGCCA
GTTACCTTCG
GAAAAAGAGT
TGGTAGCTCT
TGATCCGGCA
AACAAACCAC
CGCTGGTAGC
GGTGGTTTTT
TTGTTTGCAA
GCAGCAGATT
ACGCGCAGAA
AAAAAGGATC
TCAAGAAGAT
CCTTTGATCT
TTTCTACGGG
GTCTGACGCT
CATGGAACAA
TAAACGAAAG
GCTCAGTCGA
AAGACTGGGC
CTTTCGTTTA
TCTGTTGTTT
GAAAAAAAAA
AGCCCCGCGA
TTGCGGGGCT
TTTTTTTTTC
ATATGTCTGA
CCAGGGAAAA
TAGCCCTCTG
ACCTGGGGAT
TTGACTTCCC
TATCAGTGAT
AGAGATAATC
TGGGAGTCCC
TATCAGTGAT
AGAGAACGCG
GTATCGATAT
GTCGGACCTG
GTGCTGGGCC
TGGCCATCGG
CATCGGCTCG
GTGGGCGTGG
GCATCCTGAA
CAAGGTGACC
GGCGAGATCA
TCCACAAGAA
CTCGCGCATC
TTCCCGGCCG
CCCAGGCCGA
GAACAACCTG
GTGCGCCGCA
CCAACCGCCA
GGGCCGCCGC
CTGGCCCGCC
GCAAGAAGCA
CCGCCGCGTG
CGCCTGAACC
GCCTGTTCGA
GGAGTCGGGC
CTGATCACCG
ACTTCACCAA
GATCTCGATC
AACCTGAACC
CGTACCAGCT
GCGCGTGAAG
GGCCTGACCG
ACGAGCTGTC
GAACGAGGAG
CTGTTCATCG
CCCTGAAGAA
CATGGTGAAG
CACCGCGGCA
TCTCGTACCT
GGACGACGCC
TCGGACGACG
GCAACTCGTC
GGTGGGCGAC
TACGCCCAGA
TCGTGAAGGA
GAACTCGAAG
CAGCTGGAGA
CCAAGACCCC
GGGCCAGATC
CAGCTGGAGC
GCTACCAGAC
CTACGGCCAG
CTGCGCGGCG
ACTTCACCGT
GGAGAAGGAC
GGCAAGAAGC
ACCGCCTGAT
CAACGTGTTC
CCGACCTCGG
CCTACCGCTC
GGAGGCCCTG
CGCATCCTGC
AGACCCAGCA
GGAGTTCAAC
CCGCAGATCA
CCGACGAGTT
CATCAACCGC
TACCTGGAGA
TCCTGACCGG
CAAGCGCAAG
TACTACCACG
GCCCGGGCAA
CGAGAAGTCG
CGCACCGACT
ACGGCCGCTA
CCGCACCTCG
GGCGAGACCC
TGGACAACAT
CTTCGGCATC
CTGATCGGCA
AGTGCACCTT
CTACCCGGAC
GAGTTCCGCG
CCGCCAAGGC
CTCGTACACC
GCCCAGGAGT
TCAACCTGCT
GAACGACCTG
AACAACCTGA
CCGTGCCGAC
CGAGACCAAG
AAGCTGTCGA
AGGAGCAGAA
GAACCAGATC
ATCAACTACG
TGAAGAACGA
GAAGGCCATG
GGCCCGGCCA
AGCTGTTCAA
GTACATCGCC
AAGCTGCTGT
CGTGCGACGT
GGCCGACATC
AAGGGCTACC
GCATCGACAA
GTCGGGCAAG
GCCGAGATCC
ACACCTTCGA
GGCCTACCGC
AAGATGAAGA
CCCTGGAGAC
CCTGGACATC
GAGCAGATGG
ACCGCGAGAC
CCTGGACAAG
CTGGCCTACG
TGCTGACCCT
GAACACCGAG
CGCGAGGGCA
TCCAGGAGGC
CCTGGAGCAC
GAGTTCGCCG
ACGGCTCGTT
CTCGCAGAAG
CAGGTGGACG
AGCTGGTGCA
GTTCCGCAAG
GCCAACTCGT
CGATCTTCGG
CAAGGGCTGG
CACAACTTCT
CGGTGAAGCT
GATGATGGAG
CTGATCCCGG
AGCTGTACGA
GACCTCGGAG
GAGCAGATGA
CCATCCTGAC
CCGCCTGGGC
AAGCAGAAGA
CCACCTCGTC
GTCGAACAAG
ACCAAGTACA
TCGACGAGAA
GCTGCTGACC
GAGGAGATCT
ACAACCCGGT
GGTGGCCAAG
TCGGTGCGCC
AGGCCATCAA
GATCGTGAAC
GCCGCCATCA
AGGAGTACGG
CGACTTCGAC
AACATCGTGA
TCGAGATGGC
CCGCGAGACC
AACGAGGACG
ACGAGAAGAA
GGCCATCCAG
AAGATCCAGA
AGGCCAACAA
GGACGAGAAG
GACGCCGCCA
TGCTGAAGGC
CGCCAACCAG
TACAACGGCA
AGGCCGAGCT
GCCGCACTCG
GTGTTCCACG
GCCACAAGCA
GCTGGCCACC
AAGATCCGCC
TGTGGCACCA
GCAGGGCGAG
CGCTGCCTGT
ACACCGGCAA
GACCATCTCG
ATCCACGACC
TGATCAACAA
CTCGAACCAG
TTCGAGGTGG
ACGCCATCCT
GCCGCTGTCG
ATCACCTTCG
ACGACTCGCT
GGCCAACAAG
GTGCTGGTGT
ACGCCACCGC
CAACCAGGAG
AAGGGCCAGC
GCACCCCGTA
CCAGGCCCTG
GACTCGATGG
ACGACGCCTG
GTCGTTCCGC
GAGCTGAAGG
CCTTCGTGCG
CGAGTCGAAG
ACCCTGTCGA
ACAAGAAGAA
GGAGTACCTG
CTGACCGAGG
AGGACATCTC
GAAGTTCGAC
GTGCGCAAGA
AGTTCATCGA
GCGCAACCTG
GTGGACACCC
GCTACGCCTC
GCGCGTGGTG
CTGAACGCCC
TGCAGGAGCA
CTTCCGCGCC
CACAAGATCG
ACACCAAGGT
GTCGGTGGTG
CGCGGCCAGT
TCACCTCGCA
GCTGCGCCGC
CACTGGGGCA
TCGAGAAGAC
CCGCGACACC
TACCACCACC
ACGCCGTGGA
CGCCCTGATC
ATCGCCGCCT
CGTCGCAGCT
GAACCTGTGG
AAGAAGCAGA
AGAACACCCT
GGTGTCGTAC
TCGGAGGACC
AGCTGCTGGA
CATCGAGACC
GGCGAGCTGA
TCTCGGACGA
CGAGTACAAG
GAGTCGGTGT
TCAAGGCCCC
GTACCAGCAC
TTCGTGGACA
CCCTGAAGTC
GAAGGAGTTC
GAGGACTCGA
TCCTGTTCTC
GTACCAGGTG
GACTCGAAGT
TCAACCGCAA
GATCTCGGAC
GCCACCATCT
ACGCCACCCG
CCAGGCCAAG
GTGGGCAAGG
ACAAGGCCGA
CGAGACCTAC
GTGCTGGGCA
AGATCAAGGA
CATCTACACC
CAGGACGGCT
ACGACGCCTT
CATGAAGATC
TACAAGAAGG
ACAAGTCGAA
GTTCCTGATG
TACCGCCACG
ACCCGCAGAC
CTTCGAGAAG
GTGATCGAGC
CGATCCTGGA
GAACTACCCG
AACAAGCAGA
TCAACGACAA
GGGCAAGGAG
GTGCCGTGCA
ACCCGTTCCT
GAAGTACAAG
GAGGAGCACG
GCTACATCCG
CAAGTACTCG
AAGAAGGGCA
ACGGCCCGGA
GATCAAGTCG
CTGAAGTACT
ACGACTCGAA
GCTGGGCAAC
CACATCGACA
TCACCCCGAA
GGACTCGAAC
AACAAGGTGG
TGCTGCAGTC
GGTGTCGCCG
TGGCGCGCCG
ACGTGTACTT
CAACAAGACC
ACCGGCAAGT
ACGAGATCCT
GGGCCTGAAG
TACGCCGACC
TGCAGTTCGA
CAAGGGCACC
GGCACCTACA
AGATCTCGCA
GGAGAAGTAC
AACGACATCA
AGAAGAAGGA
GGGCGTGGAC
TCGGACTCGG
AGTTCAAGTT
CACCCTGTAC
AAGAACGACC
TGCTGCTGGT
GAAGGACACC
GAGACCAAGG
AGCAGCAGCT
GTTCCGCTTC
CTGTCGCGCA
CCATGCCGAA
GCAGAAGCAC
TACGTGGAGC
TGAAGCCGTA
CGACAAGCAG
AAGTTCGAGG
GCGGCGAGGC
CCTGATCAAG
GTGCTGGGCA
ACGTGGCCAA
CTCGGGCCAG
TGCAAGAAGG
GCCTGGGCAA
GTCGAACATC
TCGATCTACA
AGGTGCGCAC
CGACGTGCTG
GGCAACCAGC
ACATCATCAA
GAACGAGGGC
GACAAGCCGA
AGCTGGACTT
CTAAATACTC
TGGCCATTTT
TCTTTACCGC
CTTTGAGTGA
GCTGATACCG
CTCGCCGCAG
CCGAACGACC
GAGCGCAACG
CGTGCGGCCG
CTGATTAGCT
AAGCAGAAGG
CCATCCTGAC
GGATGGCCTT
TTTCTAGAAA
AAAAAAAAGC
GCCGCAACTG
CGGCGCTTTT
TTTTTTTTAA
TTAAGTGATA
GATTGCGATG
CGTGTGCTCG
GTTGATATAA
TCTGTCAGAT
CAGGAGGACT
TCGCATGTCG
CGGTTGGATA
AGTCCAAGGT
CATCAACTCG
GCGCTGGAGC
TGCTGAACGA
GGTCGGTATC
GAGGGACTGA
CCACGCGCAA
GTTGGCCCAA
AAGCTCGGCG
TGGAACAACC
GACCCTCTAC
TGGCACGTGA
AGAACAAGCG
CGCGCTGCTG
GATGCGTTGG
CCATCGAGAT
GCTGGATCGC
CATCACACGC
ACTTTTGCCC
CCTGGAGGGC
GAGAGCTGGC
AGGATTTTCT
GCGGAACAAC
GCGAAGTCGT
TCCGGTGCGC
CCTCCTGAGC
CATCGGGATG
GCGCCAAAGT
CCACTTGGGC
ACCCGCCCGA
CCGAGAAGCA
ATACGAGACC
CTGGAGAACC
AGCTGGCCTT
CCTCTGTCAA
CAGGGTTTTT
CGTTGGAGAA
CGCCCTGTAT
GCCCTGTCCG
CCGTGGGACA
CTTCACCTTG
GGCTGCGTGT
TGGAGGATCA
GGAACATCAG
GTGGCGAAAG
AGGAACGGGA
GACCCCCACG
ACGGATTCGA
TGCCCCCGTT
GTTGCGCCAG
GCGATCGAAC
TGTTTGACCA
CCAGGGCGCC
GAACCCGCCT
TTTTGTTCGG
GCTGGAGCTG
ATCATCTGCG
GGCTGGAAAA
GCAACTGAAG
TGCGAAAGCG
GTTCGTGATA
ACGCCGAAAA
CCCCGCTTCG
GCGGGGTTTT
GCCGCGGTCT
GCGTTGTCGG
GAAGATTTAA
AATAAAAAAG
GGGACCTCTA
GGGTCCCCAA
TTAAATAGTT
AACACTAGTA
TTAAAAAAAA
AAGCCCCGCG
ATTGCGGGGC
TTTTTTTTTT
CGACGGCGAG
GGTTCCGACC
GCTGCAACTC
CCGGTGCAAC
CTTGTCCCGG
TCTATTCTCT
TCACTGCACC
AGCTCCAATC
TGGTGTGAAT
GCCCCTCGTC
TGTTCGCGCA
GGCGGGGGGC
TCTATTCGTT
TGTCAGCATC
GAAAGTAGCC
AGATCAGGGA
TGCGTTGCAA
CCGCGTATGC
CCAGGTCAGA
AGAGTCGCAC
AAGAGTTGCA
GACCCCTGGA
AAGAAAAATG
GCCAGAGGGC
GAAAACACCC
TCTGACCAGC
GGAGCGGGCG
ACGGGAATCG
AACCCGCGTA
GCTAGTTTGG
AAGAATGGGT
GTCTGCCGAC
CACATATATG
GGCCGGTCAA
GATAGGTTTT
TACCCCCTCT
CGGCTGCATC
CTCTAAGTGG
AAAGAAATTG
CAGGTCGTAG
AAGCGCGTTG
AAGCCTGAGA
GTTGCACAGG
AGTTGCAACC
CGGTAGCCTT
GTTCACGACG
AGAGGAGACC
TAGTTGGCAC
GTCGCGGATG
GGGATCGCTG
AAGACTCAGC
GCAGCGGGAG
GATCCAAGCC
TCATACGTCA
ACCCGCAGGA
CGGTGTGAGG
TACTACGCGC
TGCAGACCTA
CGACAACAAG
ATGGACGCCG
AAGCCTGGCT
CGCGGGCGAG
AAGCGGCTCA
TCGAGATGGA
GACCTGGACC
CCTCCACAGG
ACCGGGCGAA
GAAGGCAGCC
GCCAGCGCCA
TCACGCTGGA
GGAGTACACC
CGGAAGTGGC
TCGTGGAGCG
CGACCTCGCA
GACGGCACCA
GGGATCTGTA
CAGCGGGCAC
GCGGAGCGCC
GCATCTACCC
GGTGCTAGGT
GAAGTGGCGG
TCACAGAGAT
GACGCCAGCT
CTGGTGCGTG
CGTGGTGGGC
CGGGATGGGT
AGGAAGCACC
CGACTGCCCG
CCGGCATGCC
TACAACGTCC
TCCGGGCGGT
GATGAACACA
GCGGTCGAGG
ACAAGCTGAT
CGCAGAGAAC
CCGTGCCGGA
TCGAGCAGAA
GGCAGCCGAT
GAGCGCGACG
TAGAGGCGCT
GACGCCTGAG
GAGCTGGACA
TCGTCGCCGC
TGAGATCTTC
GTGCACTACC
GGATCGCGGC
ATACATCCTG
GCGTGGACGA
GCCTCCGGTT
CGGAGAGCTG
ATCGAGCTTC
GCCGCAAGGA
CATCGTGGAC
GACGGCATGA
CGATGAAGCT
CCGGGTGCGC
CGTGGCGCTT
CCCGCGTGGG
GAACAAGATC
GTCGTTGGCA
ACGCCAAGAC
CGTCCGGTCG
AAGCGTCCTG
TGACGGTTCC
GCCTCACGTC
GCGGAGATGA
TCCGAGCGCA
CATGAAGGAC
CGTACGAAGA
TGAACAAGGG
CCCCGAGGCA
TTCCTGGTGA
CCACGACGCA
GGGCAACCGG
CTGTCGAAGT
CCGCGTTCAC
CAAGTCGCTG
AAGCGTGGCT
ACGCCAAGAT
CGGTCGGCCG
GAACTCCGCA
TCCACGACCT
CCGCGCTGTC
GGCGCTACGT
TCGCCGCTCA
GGCAGGTGCG
ACGACCAAGG
AGCTGATGGC
CCGCCTCGGT
CACACGACTC
CTAGGATGGC
GATGAAGTAC
CAGATGGCGT
CTGAGGCCCG
CGACGAGGCT
ATCGCTGAGG
CGATGTCCAA
GCTGGCCAAG
ACCTCCTGAA
ACGCAAAAAG
CCCCCCTCCC
AAGGACACTG
AGTCCTAAAG
AGGGGGGTTT
CTTGTCAGTA
CGCGAAGAAC
CACGCCTGGC
CGCGAGCGCC
AGCACCGCCG
CTCTGTGCGG
AGACCTGGGC
ACCAGCCCCG
CCGCCGCCAG
GAGCATTGCC
GTTCCCGCCA
GCTAGCCACC
TGACGTCGGG
GGGGGGGGAA
AGCCACGTTG
TGTCTCAAAA
TCTCTGATGT
TACATTGCAC
AAGATAAAAA
TATATCATCA
TGAACAATAA
AACTGTCTGC
TTACATAAAC
AGTAATACAA
GGGGTGTTAT
GAGCCATATT
CAACGGGAAA
CGTCTTGCTC
GAGGCCGCGA
TTAAATTCCA
ACATGGATGC
TGATTTATAT
GGGTATAAAT
GGGCTCGCGA
TAATGTCGGG
CAATCAGGTG
CGACAATCTA
TCGCTTGTAT
GGGAAGCCCC
ATGCGCCAGA
GTTGTTTCTG
AAACATGGCA
AAGGTAGCGT
TGCCAATGAT
GTTACAGATG
AGATGGTCAG
ACTAAACTGG
CTGACGGAAT
TTATGCCTCT
TCCGACCATC
AAGCATTTTA
TCCGTACTCC
TGATGATGCA
TGGTTACTCA
CCACTGCGAT
CCCCGGGAAA
ACAGCATTCC
AGGTATTAGA
AGAATATCCT
GATTCAGGTG
AAAATATTGT
TGATGCGCTG
GCAGTGTTCC
TGCGCCGGTT
GCATTCGATT
CCTGTTTGTA
ATTGTCCTTT
TAACAGCGAT
CGCGTATTTC
GCCTCGCTCA
GGCGCAATCA
CGAATGAATA
ACGGTTTGGT
TGATGCGAGT
GATTTTGATG
ACGAGCGTAA
TGGCTGGCCT
GTTGAACAAG
TCTGGAAAGA
AATGCATAAT
CTTTTGCCAT
TCTCACCGGA
TTCAGTCGTC
ACTCATGGTG
ATTTCTCACT
TGATAACCTT
ATTTTTGACG
AGGGGAAATT
AATAGGTTGT
ATTGATGTTG
GACGAGTCGG
AATCGCAGAC
CGATACCAGG
ATCTTGCCAT
CCTATGGAAC
TGCCTCGGTG
AGTTTTCTCC
TTCATTACAG
AAACGGCTTT
TTCAAAAATA
TGGTATTGAT
AATCCTGATA
TGAATAAATT
GCAGTTTCAT
TTGATGCTCG
ATGAGTTTTT
CTAATCAGAA
TTGGTTAATT
GGTTGTAACA
CTGGCAGAGC
ATTACGCTGA
CTTGACGGGA
CGGCGGCTTT
GTTGAATAAA
TCGAACTTTT
GCTGAGTTGA
AGGATCAGAT
CACGCATCTT
CCCGACAACG
CAGACCGTTC
CGTGGCAAAG
CAAAAGTTCA
AAATCACCAA
CTGGTCCACC
TACAACAAAG
CTCTCACCAA
CCGTGGCTCC
CTCACGATAT
C