名称: | pHY-P43 |
---|---|
别称: | |
基因长: | |
质粒宿主: | 枯草杆菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 空载体 |
原核抗性: | Amp |
筛选标记: | Tet |
荧光标记: | |
启动子: | p43 |
复制子: | repB |
拷贝数: | |
感受态: | DH5a |
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。
AAGCTTTGAT
AGGTGGTATG
TTTTCGCTTG
AACTTTTAAA
TACAGCCATT
GAACATACGG
TTGATTTAAT
AACTGACAAA
CATCACCCTC
TTGCTAAAGC
GGCCAAGGAC
GCTGCCGCCG
GGGCTGTTTG
CGTTTTTGCC
GTGATTTCGT
GTATCATTGG
TTTACTTATT
TTTTTGCCAA
AGCTGTAATG
GCTGAAAATT
CTTACATTTA
TTTTACATTT
TTAGAAATGG
GCGTGAAAAA
AAGCGCGCGA
TTATGTAAAA
TATAAAGTGA
TAGCGGTACC
ATTATAGGTA
AGAGAGGAAC
GTACACAGAT
CTGCAGGTCG
ACGGATCCCC
GGGAATTCCT
GTTATAAAAA
AAGGATCAAT
TTTGAACTCT
CTCCCAAAGT
TGATCCCTTA
ACGATTTAGA
AATCCCTTTG
AGAATGTTTA
TATACATTCA
AGGTAACCAG
CCAACTAATG
ACAATGATTC
CTGAAAAAAG
TAATAACAAA
TTACTATACA
GATAAGTTGA
CTGATCAACT
TCCATAGGTA
ACAACCTTTG
ATCAAGTAAG
GGTATGGATA
ATAAACCACC
TACAATTGCA
ATACCTGTTC
CCTCTGATAA
AAAGCTGGTA
AAGTTAAGCA
AACTCATTCC
AGCACCAGCT
TCCTGCTGTT
TCAAGCTACT
TGAAACAATT
GTTGATATAA
CTGTTTTGGT
GAACGAAAGC
CCACCTAAAA
CAAATACGAT
TATAATTGTC
ATGAACCATG
ATGTTGTTTC
TAAAAGAAAG
GAAGCAGTTA
AAAAGCTAAC
AGAAAGAAAT
GTAACTCCGA
TGTTTAACAC
GTATAAAGGA
CCTCTTCTAT
CAACAAGTAT
CCCACCAATG
TAGCCGAAAA
TAATGACACT
CATTGTTCCA
GGGAAAATAA
TTACACTTCC
GATTTCGGCA
GTACTTAGCT
GGTGAACATC
TTTCATCATA
TAAGGAACCA
TAGAGACAAA
CCCTGCTACT
GTTCCAAATA
TAATTCCCCC
ACAAAGAACT
CCAATCATAA
AAGGTATATT
TTTCCCTAAT
CCGGGATCAA
CAAAAGGATC
TGTTACTTTC
CTGATATGTT
TTACAAATAT
CAGGAATGAC
AGCACGCTAA
CGATAAGAAA
AGAAATGCTA
TATGATGTTG
TAAACAACAT
AAAAAATACA
ATGCCTACAG
ACATTAGTAT
AATTCCTTTG
ATATCAAAAT
GACCTTTTAT
CCTTACTTCT
TTCTTTAATA
ATTTCATAAG
AAACGGAACA
GTGATAATTG
TTATCATAGG
AATGAGTAGA
AGATAGGACC
AATGAATATA
ATGGGCTATC
ATTCCACCAA
TCGCTGGACC
GACTCCTTCT
CCCATGGCTA
CTATCGATCC
AATAAGACCA
AATGCTTTAC
CCCTATTTTC
CTTTGGAATA
TAGCGCGCAA
CTACAACCAT
TACGAGTGCT
GGAAATGCAG
CTGCACCAGC
CCCTTGAATA
AAACGAGCCA
TAATAAGTAA
GGAAAAGAAA
GAATGGCCAA
CAAACCCAAT
TACCGACCCG
AAACAATTTA
TTATAATTCC
AAATAGGAGT
AACCTTTTGA
TGCCTAATTG
ATCAGATAGC
TTTCCATATA
CAGCTGTTCC
AATGGAAAAG
GTTAACATAA
AGGCTGTGTT
CACCCAGTTT
GTACTCGCAG
GTGGTTTATT
AAAATCATTT
GCAATATCAG
GTAATGAGAC
GTTCAAAACC
ATTTCATTTA
ATACGCTAAA
AAAAGATAAA
ATGCAAAGCC
AAATTAAAAT
TTGGTTGTGT
CGTAAATTCG
ATTGTGAATA
GGATGTATTC
ACATTTCACC
CTCCAATAAT
GAGGGCAGAC
GTAGTTTATA
GGGTTAATGA
TACGCTTCCC
TCTTTTAATT
GAACCCTGTT
ACATTCATTA
TTCATTACAC
TTCATAATTA
ATTCCTCCTA
AACTTGATTA
AAACATTTTA
CCACATATAA
ACTAAGTTTT
AAATTCAGTA
TTTCATCACT
TATACAACAA
TATGGCCCGT
TTGTTGAACT
ACTCTTTAAT
AAAATAATTT
TTCCGTTCCC
AATTCCACAT
TGCAATAATA
GAAAATCCAT
CTTCATCGGC
TTTTTCGTCA
TCATCTGTAT
GAATCAAATC
GCCTTCTTCT
GTGTCATCAA
GGTTTAATTT
TTTATGTATT
TCTTTTAACA
AACCACCATA
GGAGATTAAC
CTTTTACGGT
GTAAACCTTC
CTCCAAATCA
GACAAACGTT
TCAAATTCTT
TTCTTCATCA
TCGGTCATAA
AATCCGTATC
CTTTACAGGA
TATTTTGCAG
TTTCGTCAAT
TGCCGATTGT
ATATCCGATT
TATATTTATT
TTTCGGTCGA
ATCATTTGAA
CTTTTACATT
TGGATCATAG
TCTAATTTCA
TTGCCTTTTT
CCAAAATTGA
ATCCATTGTT
TTTGATTCAC
GTAGTTTTCT
GTATTCTTAA
AATAAGTTGG
TTCCACACAT
ACCAATACAT
GCATGTGCTG
ATTATAAGAA
TTATCTTTAT
TATTTATTGT
CACTTCCGTT
GCACGCATAA
AACCAACAAG
ATTTTTATTA
ATTTTTTTAT
ATTGCATCAT
TCGGCGAAAT
CCTTGAGCCA
TATCTGACAA
ACTCTTATTT
AATTCTTCGC
CATCATAAAC
ATTTTTAACT
GTTAATGTGA
GAAACAACCA
ACGAACTGTT
GGCTTTTGTT
TAATAACTTC
AGCAACAACC
TTTTGTGACT
GAATGCCATG
TTTCATTGCT
CTCCTCCAGT
TGCACATTGG
ACAAAGCCTG
GATTTACAAA
ACCACACTCG
ATACAACTTT
CTTTCGCCTG
TTTCACGATT
TTGTTTATAC
TCTAATATTT
CAGCACAATC
TTTTACTCTT
TCAGCCTTTT
TAAATTCAAG
AATATGCAGA
AGTTCAAAGT
AATCAACATT
AGCGATTTTC
TTTTCTCTCC
ATGGTCTCAC
TTTTCCACTT
TTTGTCTTGT
CCACTAAAAC
CCTTGATTTT
TCATCTGAAT
AAATGCTACT
ATTAGGACAC
ATAATATTAA
AAGAAACCCC
CATCTATTTA
GTTATTTGTT
TGGTCACTTA
TAACTTTAAC
AGATGGGGTT
TTTCTGTGCA
ACCAATTTTA
AGGGTTTTCC
AATACTTTAA
AACACATACA
TACCAACACT
TCAACGCACC
TTTCAGCAAC
TAAAATAAAA
ATGACGTTAT
TTCTATATGT
ATCAAGATAA
GAAAGAACAA
GTTCAAAACC
ATCAAAAAAA
GACACCTTTT
CAGGTGCTTT
TTTTATTTTA
TAAACTCATT
CCCTGATCTC
GACTTCGTTC
TTTTTTTACC
TCTCGGTTAT
GAGTTAGTTC
AAATTCGTTC
TTTTTAGGTT
CTAAATCGTG
TTTTTCTTGG
AATTGTGCTG
TTTTATCCTT
TACCTTGTCT
ACAAACCCCT
TAAAAACGTT
TTTAAAGGCT
TTTAAGCGTC
TGTACGTTCC
TTAAGGAATT
ATTCCTTAGT
GCTTTCTAGG
TTAATGTCAT
GATAATAATG
GTTTCTTAGA
CGTCAGGTGG
CACTTTTCGG
GGAAATGTCC
GCGGAACCCC
TATTTGTTTA
TTTTTCTAAA
TACATTCAAA
TATGTATCCG
CTCATGAGAC
AATAACCCTG
ATAAATGCTT
CAATAATATT
GAAAAAGGAA
GAGTATGAGT
ATTCAACATT
TCCGTGTCGC
CCTTATTCCC
TTTTTTGCGG
CATTTTGCCT
TCCTGTTTTT
GCTCACCCAG
AAACGCTGGT
GAAAGTAAAA
GATGCTGAAG
ATCAGTTGGG
TGCACGAGTG
GGTTACATCG
AACTGGATCT
CAACAGCGGT
AAGATCCTTG
AGAGTTTTCG
CCCCGAAGAA
CGTTTTCCAA
TGATGAGCAC
TTTTAAAGTT
CTGCTATGTG
GCGCGGTATT
ATCCCGTGTT
GACGCCGGGC
AAGAGCAACT
CGGTCGCCGC
ATACACTATT
CTCAGAATGA
CTTGGTTGAG
TACTCACCAG
TCACAGAAAA
GCATCTTACG
GATGGCATGA
CAGTAAGAGA
ATTATGCAGT
GCTGCCATAA
CCATGAGTGA
TAACACTGCG
GCCAACTTAC
TTCTGACAAC
GATCGGAGGA
CCGAAGGAGC
TAACCGCTTT
TTTGCACAAC
ATGGGGGATC
ATGTAACTCG
CCTTGATCGT
TGGGAACCGG
AGCTGAATGA
AGCCATACCA
AACGACGAGC
GTGACACCAC
GATGCCTGCA
GCAATGGCAA
CAACGTTGCG
CAAACTATTA
ACTGGCGAAC
TACTTACTCT
AGCTTCCCGG
CAACAATTAA
TAGACTGGAT
GGAGGCGGAT
AAAGTTGCAG
GACCACTTCT
GCGCTCGGCC
CTTCCGGCTG
GCTGGTTTAT
TGCTGATAAA
TCTGGAGCCG
GTGAGCGTGG
GTCTCGCGGT
ATCATTGCAG
CACTGGGGCC
AGATGGTAAG
CCCTCCCGTA
TCGTAGTTAT
CTACACGACG
GGGAGTCAGG
CAACTATGGA
TGAACGAAAT
AGACAGATCG
CTGAGATAGG
TGCCTCACTG
ATTAAGCATT
GGTAACTGTC
AGACCAAGTT
TACTCATATA
TACTTTAGAT
TGATTTAAAA
CTTCATTTTT
AATTTAAAAG
GATCTAGGTG
AAGATCCTTT
TTGATAATCT
CATGACCAAA
ATCCCTTAAC
GTGAGTTTTC
GTTCCACTGA
GCGTCAGACC
CCTTAATAAG
ATGATCTTCT
TGAGATCGTT
TTGGTCTGCG
CGTAATCTCT
TGCTCTGAAA
ACGAAAAAAC
CGCCTTGCAG
GGAGGTTTTT
CGAAGGTTCT
CTGAGCTACC
AACTCTTTGA
ACCGAGGTAA
CTGGCTTGCA
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
GTCAGGGGGC
GGAGCCTATG
GAAAAACGCT
TTGCCC