名称: | pFLP-Hyg |
---|---|
别称: | Plasmid#87831 |
基因长: | |
质粒宿主: | 大肠杆菌 |
质粒用途: | 基因编辑 |
片段类型: | |
片段物种: | |
原核抗性: | Hyg |
筛选标记: | |
荧光标记: | |
启动子: | |
复制子: | p15A |
拷贝数: | 低 |
感受态: | DH5a |
温度: | 30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。
ATGATCTTCT
TGAGATCGTT
TTGGTCTGCG
CGTAATCTCT
TGCTCTGAAA
ACGAAAAAAC
CGCCTTGCAG
GGCGGTTTTT
CGAAGGTTCT
CTGAGCTACC
AACTCTTTGA
ACCGAGGTAA
CTGCCTTGGA
GGAGCGCAGT
CACCAAAACT
TGTCCTTTCA
GTTTAGCCTT
AACCGGCGCA
TGACTTCAAG
ACTAACTCCT
CTAAATCAAT
TACCAGTGGC
TGCTGCCAGT
GGTGCTTTTG
CATGTCTTTC
CGGGTTGGAC
TCAAGACGAT
AGTTACCGGA
TAAGGCGCAG
CGGTCGGACT
GAACGGGGGG
TTCGTGCATA
CAGTCCAGCT
TGGAGCGAAC
TGCCTACCCG
GAACTGAGTG
TCAGGCGTGG
AATGAGACAA
ACGCGGCCAT
AACAGCGGAA
TGACACCGGT
AAACCGAAAG
GCAGGAACAG
GAGAGCGCAC
GAGGGAGCCG
CCAGGGGGAA
ACGCCTGGTA
TCTTTATAGT
CCTGTCGGGT
TTCGCCACCA
CTGATTTGAG
CGTCAGATTT
CGTGATGCTT
GTCAGGGGGG
CGGAGCCTAT
GGAAAAACGG
CTTTGCCGCG
GCCCTCTCAC
TTCCCTGTTA
AGTATCTTCC
TGGCATCTTC
CAGGAAATCT
CCGCCCCGTT
CGTAAGCCAT
TTCCGCTCGC
CGCAGTCGAA
CGACCGAGCG
TAGCGAGTCA
GTGAGCGAGG
AAGCGGAATA
TATCCTGTAT
CACATATTCT
GCTGACGCAC
CGGTGCAGCC
TTTTTTCTCC
TGCCACATGA
AGCACTTCAC
TGACACCCTC
ATCAGTGCCA
ACATAGTAAG
CCAGTATACA
CTCCGCTAGC
GCTGATGTCC
GGCGGTGCTT
TTGCCGTTAC
GCACCACCCC
GTCAGTAGCT
GAACAGGAGG
GACAGCTGAT
AGAAACAGAA
GCCACTGGAG
CACCTCAAAA
ACACCATCAT
ACACTAAATC
AGTAAGTTGG
CAGCATCACC
CGACGCACTT
TGCAGATCTC
GGGGAAATGT
GCGCGGAACC
CCTATTTGTT
TATTTTTCTA
AATACATTCA
AATATGTATC
CGCTCATGAG
ACAATAACCC
TGATAAATGC
TTCAATAATA
TTGAAAAAGG
AAGAGTATGA
AAAAGCCTGA
ACTCACCGCG
ACGTCTGTCG
AGAAGTTTCT
GATCGAAAAG
TTCGACAGCG
TCTCCGACCT
GATGCAGCTC
TCGGAGGGCG
AAGAATCTCG
TGCTTTCAGC
TTCGATGTAG
GAGGGCGTGG
ATATGTCCTG
CGGGTAAATA
GCTGCGCCGA
TGGTTTCTAC
AAAGATCGTT
ATGTTTATCG
GCACTTTGCA
TCGGCCGCGC
TCCCGATTCC
GGAAGTGCTT
GACATTGGGG
AATTCAGCGA
GAGCCTGACC
TATTGCATCT
CCCGCCGTGC
ACAGGGTGTC
ACGTTGCAAG
ACCTGCCTGA
AACCGAACTG
CCCGCTGTTC
TGCAGCCGGT
CGCGGAGGCC
ATGGATGCGA
TCGCTGCGGC
CGATCTTAGC
CAGACGAGCG
GGTTCGGCCC
ATTCGGACCG
CAAGGAATCG
GTCAATACAC
TACATGGCGT
GATTTCATAT
GCGCGATTGC
TGATCCCCAT
GTGTATCACT
GGCAAACTGT
GATGGACGAC
ACCGTCAGTG
CGTCCGTCGC
GCAGGCTCTC
GATGAGCTGA
TGCTTTGGGC
CGAGGACTGC
CCCGAAGTCC
GGCACCTCGT
GCACGCGGAT
TTCGGCTCCA
ACAATGTCCT
GACGGACAAT
GGCCGCATAA
CAGCGGTCAT
TGACTGGAGC
GAGGCGATGT
TCGGGGATTC
CCAATACGAG
GTCGCCAACA
TCTTCTTCTG
GAGGCCGTGG
TTGGCTTGTA
TGGAGCAGCA
GACGCGCTAC
TTCGAGCGGA
GGCATCCGGA
GCTTGCAGGA
TCGCCGCGGC
TCCGGGCGTA
TATGCTCCGC
ATTGGTCTTG
ACCAACTCTA
TCAGAGCTTG
GTTGACGGCA
ATTTCGATGA
TGCAGCTTGG
GCGCAGGGTC
GATGCGACGC
AATCGTCCGA
TCCGGAGCCG
GGACTGTCGG
GCGTACACAA
ATCGCCCGCA
GAAGCGCGGC
CGTCTGGACC
GATGGCTGTG
TAGAAGTACT
CGCCGATAGT
GGAAACCGAC
GCCCCAGCAC
TCGTCCGAGG
GCAAAGGAAT
AGCGTTCTTC
GGGGCGAAAA
CTCTCAAGGA
TCTTACCGCT
GTTGAGATCC
AGTTCGATGT
AACCCACTCG
TGCACCCAAC
TGATCTTCAG
CATCTTTTAC
TTTCACCAGC
GTTTCTGGGT
GAGCAAAAAC
AGGAAGGCAA
AATGCCGCAA
AAAAGGGAAT
AAGGGCGACA
CGGAAATGTT
GAATACTCAT
ACTCTTCCTT
TTTCAATATT
ATTGAAGCAT
TTATCAGGGT
TATTGTCTCA
TGAGCGGATA
CATATTTGAA
TGTATTTAGA
AAAATAAACA
AATAGGGGTT
CCGCGCACAT
TTCCCCGAAA
AGTGCCACCT
GACGTCTAAG
AAACCATTAT
TATCATGACA
TTAACCTATA
AAAATAGGCG
TATCACGAGG
CCCTTTCGTC
TTCAAGAATT
TTATAAACCG
TGGAGCGGGC
AATACTGAGC
TGATGAGCAA
TTTCCGTTGC
ACCAGTGCCC
TTCTGATGAA
GCGTCAGCAC
GACGTTCCTG
TCCACGGTAC
GCCTGCGGCC
AAATTTGATT
CCTTTCAGCT
TTGCTTCCTG
TCGGCCCTCA
TTCGTGCGCT
CTAGGATCCT
CTACGCCGGA
CGCATCGTGG
CCGGCATCAC
CGGCGCTGAG
GTCTGCCTCG
TGAAGAAGGT
GTTGCTGACT
CATACCAGGC
CTGAATCGCC
CCATCATCCA
GCCAGAAAGT
GAGGGAGCCA
CGGTTGATGA
GAGCTTTGTT
GTAGGTGGAC
CAGTTGGTGA
TTTTGAACTT
TTGCTTTGCC
ACGGAACGGT
CTGCGTTGTC
GGGAAGATGC
GTGATCTGAT
CCTTCAACTC
AGCAAAAGTT
CGATTTATTC
AACAAAGCCG
CCGTCCCGTC
AAGTCAGCGT
AATGCTCTGC
CAGTGTTACA
ACCAATTAAC
CAATTCTGAT
TAGAAAAACT
CATCGAGCAT
CAAATGAAAC
TGCAATTTAT
TCATATCAGG
ATTATCAATA
CCATATTTTT
GAAAAAGCCG
TTTCTGTAAT
GAAGGAGAAA
ACTCACCGAG
GCAGTTCCAT
AGGATGGCAA
GATCCTGGTA
TCGGTCTGCG
ATTCCGACTC
GTCCAACATC
AATACAACCT
ATTAATTTCC
CCTCGTCAAA
AATAAGGTTA
TCAAGTGAGA
AATCACCATG
AGTGACGACT
GAATCCGGTG
AGAATGGCAG
AATAGGAACT
TCGGAATAGG
AACTTCAAAG
CGTTTCCGAA
AACGAGCGCT
TCCGAAAATG
CAACGCGAGC
TGCGCACATA
CAGCTCACTG
TTCACGTCGC
ACCTATATCT
GCGTGTTGCC
TGTATATATA
TATACATGAG
AAGAACGGCA
TAGTGCGTGT
TTATGCTTAA
ATGCGTACTT
ATATGCGTCT
ATTTATGTAG
GATGAAAGGT
AGTCTAGTAC
CTCCTGTGAT
ATTATCCCAT
TCCATGCGGG
GTATCGTATG
CTTCCTTCAG
CACTACCCTT
TAGCTGTTCT
ATATGCTGCC
ACTCCTCAAT
TGGATTAGTC
TCATCCTTCA
ATGCTATCAT
TTCCTTTGAT
ATTGGATCAT
ATGCATAGTA
CCGAGAAACT
AGTGCGAAGT
AGTGATCAGG
TATTGCTGTT
ATCTGATGAG
TATACGTTGT
CCTGGCCACG
GCAGAAGCAC
GCTTATCGCT
CCAATTTCCC
ACAACATTAG
TCAACTCCGT
TAGGCCCTTC
ATTGAAAGAA
ATGAGGTCAT
CAAATGTCTT
CCAATGTGAG
ATTTTGGGCC
ATTTTTTATA
GCAAAGATTG
AATAAGGCGC
ATTTTTCTTC
AAAGCTTTAT
TGTACGATCT
GACTAAGTTA
TCTTTTAATA
ATTGGTATTC
CTGTTTATTG
CTTGAAGAAT
TGCCGGTCCT
ATTTACTCGT
TTTAGGACTG
GTTCAGAATT
CCTCAAAAAT
TCATCCAAAT
ATACAAGTGG
ATCGATCCTA
CCCCTTGCGC
TAAAGAAGTA
TATGTGCCTA
CTAACGCTTG
TCTTTGTCTC
TGTCACTAAA
CACTGGATTA
TTACTCCCAG
ATACTTATTT
TGGACTAATT
TAAATGATTT
CGGATCAACG
TTCTTAATAT
CGCTGAATCT
TCCACAATTG
ATGAAAGTAG
CTAGGAAGAG
GAATTGGTAT
AAAGTTTTTG
TTTTTGTAAA
TCTCGAAGTA
TACTCAAACG
AATTTAGTAT
TTTCTCAGTG
ATCTCCCAGA
TGCTTTCACC
CTCACTTAGA
AGTGCTTTAA
GCATTTTTTT
ACTGTGGCTA
TTTCCCTTAT
CTGCTTCTTC
CGATGATTCG
AACTGTAATT
GCAAACTACT
TACAATATCA
GTGATATCAG
ATTGATGTTT
TTGTCCATAG
TAAGGAATAA
TTGTAAATTC
CCAAGCAGGA
ATCAATTTCT
TTAATGAGGC
TTCCAGAATT
GTTGCTTTTT
GCGTCTTGTA
TTTAAACTGG
AGTGATTTAT
TGACAATATC
GAAACTCAGC
GAATTGCTTA
TGATAGTATT
ATAGCTCATG
AATGTGGCTC
TCTTGATTGC
TGTTCCGTTA
TGTGTAATCA
TCCAACATAA
ATAGGTTAGT
TCAGCAGCAC
ATAATGCTAT
TTTCTCACCT
GAAGGTCTTT
CAAACCTTTC
CACAAACTGA
CGAACAAGCA
CCTTAGGTGG
TGTTTTACAT
AATATATCAA
ATTGTGGCAT
ACAACCTCCT
TAGTACATGC
AACCATTATC
ACCGCCAGAG
GTAAAATAGT
CAACACGCAC
GGTGTTAGAT
ATTTATCCCT
TGCGGTGATA
GATTTAACGT
ATGAGCACAA
AAAAGAAACC
ATTAACACAA
GAGCAGCTTG
AGGACGCACG
TCGCCTTAAA
GCAATTTATG
AAAAAAAGAA
AAATGAACTT
GGCTTATCCC
AGGAATCTGT
CGCAGACAAG
ATGGGGATGG
GGCAGTCAGG
CGTTGGTGCT
TTATTTAATG
GCATCAATGC
ATTAAATGCT
TATAACGCCG
CATTGCTTAC
AAAAATTCTC
AAAGTTAGCG
TTGAAGAATT
TAGCCCTTCA
ATCGCCAGAG
AAATCTACGA
GATGTATGAA
GCGGTTAGTA
TGCAGCCGTC
ACTTAGAAGT
GAGTATGAGT
ACCCTGTTTT
TTCTCATGTT
CAGGCAGGGA
TGTTCTCACC
TAAGCTTAGA
ACCTTTACCA
AAGGTGATGC
GGAGAGATGG
GTAAGCACAA
CCAAAAAAGC
CAGTGATTCT
GCATTCTGGC
TTGAGGTTGA
AGGTAATTCC
ATGACCGCAC
CAACAGGCTC
CAAGCCAAGC
TTTCCTGACG
GAATGTTAAT
TCTCGTTGAC
CCTGAGCAGG
CTGTTGAGCC
AGGTGATTTC
TGCATAGCCA
GACTTGGGGG
TGATGAGTTT
ACCTTCAAGA
AACTGATCAG
GGATAGCGGT
CAGGTGTTTT
TACAACCACT
AAACCCACAG
TACCCAATGA
TCCCATGCAA
TGAGAGTTGT
TCCGTTGTGG
GGAAAGTTAT
CGCTAGTCAG
TGGCCTGAAG
AGACGTTTGG
CTGATCGGCA
AGGTGTTCTG
GTCGGCGCAT
AGCTGATAAC
AATTGAGCAA
GAATCTGCAT
TTCTTTCCAG
ACTTGTTCAA
CAGGCCAGCC
ATTACGCTCG
TCATCAAAAT
CACTCGCATC
AACCAAACCG
TTATTCATTC
GTGATTGCGC
CTGAGCGAGA
CGAAATACGC
GATCGCTGTT
AAAAGGACAA
TTACAAACAG
GAATCGAATG
CAACCGGCGC
AGGAACACTG
CCAGCGCATC
AACAATATTT
TCACCTGAAT
CAGGATATTC
TTCTAATACC
TGGAATGCTG
TTTTCCCGGG
GATCGCAGTG
GTGAGTAACC
ATGCATCATC
AGGAGTACGG
ATAAAATGCT
TGATGGTCGG
AAGAGGCATA
AATTCCGTCA
GCCAGTTTAG
TCTGACCATC
TCATCTGTAA
CATCATTGGC
AACGCTACCT
TTGCCATGTT
TCAGAAACAA
CTCTGGCGCA
TCGGGCTTCC
CATACAATCG
ATAGATTGTC
GCACCTGATT
GCCCGACATT
ATCGCGAGCC
CATTTATACC
CATATAAATC
AGCATCCATG
TTGGAATTTA
ATCGCGGCCT
CGAGCAAGAC
GTTTCCCGTT
GAATATGGCT
CATAACACCC
CTTGTATTAC
TGTTTATGTA
AGCAGACAGT
TTTATTGTTC
ATGATGATAT
ATTTTTATCT
TGTGCAATGT
AACATCAGAG
ATTTTGAGAC
ACAACGTGGC
TTTGTTGAAT
AAATCGAACT
TTTGCTGAGT
TGAAGGATCA
GATCACGCAT
CTTCCCGACA
ACGCAGACCG
TTCCGTGGCA
AAGCAAAAGT
TCAAAATCAC
CAACTGGTCC
ACCTACAACA
AAGCTCTCAT
CAACCGTGGC
TCCCTCACTT
TCTGGCTGGA
TGATGGGGCG
ATTCAGGCCT
GGTATGAGTC
AGCAACACCT
TCTTCACGAG
GCAGACCTCA
GCGCCACAGG
TGCGGTTGCT
GGCGCTAACC
GTTTTTATCA
GGCTCTGGGA
GGCAGAATAA
ATGATCATAT
CGTCAATTAT
TACCTCCACG
GGGAGAGCCT
GAGCAAACTG
GCCTCAGGCA
TTTGAGAAGC
ACACGGTCAC
ACTGCTTCCG
GTAGTCAATA
AACCGGTAAA
CCAGCAATAG
ACATAAGCGG
CTATTTAACG
ACCCTGCCCT
GAACCGACGA
CCGGGTCGAA
TTTGCTTTCG
AATTTCTGCC
ATTCATCCGC
TTATTATCAC
TTATTCAGGC
GTAGCAACCA
GGCGTTTAAG
GGCACCAATA
ACTGCCTTAA
AAAAATTACG
CCCCGCCCTG
CCACTCATCG
CAGTACTGTT
GTAATTCATT
AAGCATTCTG
CCGACATGGA
AGCCATCACA
AACGGCATGA
TGAACCTGAA
TCGCCAGCGG
CATCAGCACC
TTGTCGCCTT
GCGTATAATA
TTTGCCCATG
GTGCACCGAT
GATAAGCTGT
CAAACATGAG
AATTACAACT
TATATCGTAT
GGGGCTGACT
TCAGGTGCTA
CATTTGAAGA
GATAAATTGC
ACTGAAATCT
AGAAATATTT
TATCTGATTA
ATAAG