名称: | pTRKH2 |
---|---|
别称: | Plasmid#71312 |
基因长: | |
质粒宿主: | 革兰氏阳性菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | |
原核抗性: | Ery |
筛选标记: | |
荧光标记: | |
启动子: | |
复制子: | p15A |
拷贝数: | 低 |
感受态: | MC1061 |
温度: | 37度 |
背景载体: | pIL253 |
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。
CCAATACGCA
AACCGCCTCT
CCCCGCGCGT
TGGCCGATTC
ATTAATGCAG
CTGGCACGAC
AGGTTTCCCG
ACTGGAAAGC
GGGCAGTGAG
CGCAACGCAA
TTAATGTGAG
TTAGCTCACT
CATTAGGCAC
CCCAGGCTTT
ACACTTTATG
CTTCCGGCTC
GTATGTTGTG
TGGAATTGTG
AGCGGATAAC
AATTTCACAC
AGGAAACAGC
TATGACCATG
ATTACGAATT
CATCGATATC
TAGATCTCGA
GCTCGCGAAA
GCTTGGCTGC
AGGTCGACGG
ATCCCCGGGA
ATTCACTGGC
CGTCGTTTTA
CAACGTCGTG
ACTGGGAAAA
CCCTGGCGTT
ACCCAACTTA
ATCGCCTTGC
AGCACATCCC
CCTTTCGCCA
GCTGGCGTAA
TAGCGAAGAG
GCCCGCACCG
ATCGCCCTTC
CCAACAGTTG
CGCAGCCTGA
ATGGCGAATG
CTAGAATCGA
TACGATTTTG
AAGTGGCAAC
AGATAAAAAA
AAGCAGTTTA
AAATTGTTGC
TGAACTTTTA
AAACAAGCAA
ATACAATCAT
TGTCGCAACA
GATAGCGACA
GAGAAGGCGA
AAACATTGCC
TGGTCGATCA
TTCATAAAGC
AAATGCCTTT
TCTAAAGATA
AAACGTATAA
AAGACTATGG
ATCAATAGTT
TAGAAAAAGA
TGTGATCCGT
AGCGGTTTTC
AAAATTTGCA
ACCAGGAATG
AATTACTATC
CCTTTTATCA
AGAAGCGCAA
AAGAAAAACG
AAATGATACA
CCAATCAGTG
CAAAAAAAGA
TATAATGGGA
GATAAGACGG
TTCGTGTTCG
TGCTGACTTG
CACCATATCA
TAAAAATCGA
AACAGCAAAG
AATGGCGGAA
ACGTAAAAGA
AGTTATGGAA
ATAAGACTTA
GAAGCAAACT
TAAGAGTGTG
TTGATAGTGC
AGTATCTTAA
AATTTTGTAT
AATAGGAATT
GAAGTTAAAT
TAGATGCTAA
AAATTTGTAA
TTAAGAAGGA
GTGATTACAT
GAACAAAAAT
ATAAAATATT
CTCAAAACTT
TTTAACGAGT
GAAAAAGTAC
TCAACCAAAT
AATAAAACAA
TTGAATTTAA
AAGAAACCGA
TACCGTTTAC
GAAATTGGAA
CAGGTAAAGG
GCATTTAACG
ACGAAACTGG
CTAAAATAAG
TAAACAGGTA
ACGTCTATTG
AATTAGACAG
TCATCTATTC
AACTTATCGT
CAGAAAAATT
AAAACTGAAT
ACTCGTGTCA
CTTTAATTCA
CCAAGATATT
CTACAGTTTC
AATTCCCTAA
CAAACAGAGG
TATAAAATTG
TTGGGAGTAT
TCCTTACCAT
TTAAGCACAC
AAATTATTAA
AAAAGTGGTT
TTTGAAAGCC
ATGCGTCTGA
CATCTATCTG
ATTGTTGAAG
AAGGATTCTA
CAAGCGTACC
TTGGATATTC
ACCGAACACT
AGGGTTGCTC
TTGCACACTC
AAGTCTCGAT
TCAGCAATTG
CTTAAGCTGC
CAGCGGAATG
CTTTCATCCT
AAACCAAAAG
TAAACAGTGT
CTTAATAAAA
CTTACCCGCC
ATACCACAGA
TGTTCCAGAT
AAATATTGGA
AGCTATATAC
GTACTTTGTT
TCAAAATGGG
TCAATCGAGA
ATATCGTCAA
CTGTTTACTA
AAAATCAGTT
TCATCAAGCA
ATGAAACACG
CCAAAGTAAA
CAATTTAAGT
ACCGTTACTT
ATGAGCAAGT
ATTGTCTATT
TTTAATAGTT
ATCTATTATT
TAACGGGAGG
AAATAATTCT
ATGAGTCGCT
TTTGTAAATT
TGGAAAGTTA
CACGTTACTA
AAGGGAATGT
AGATAAATTA
TTAGGTATAC
TACTGACAGC
TTCCAAGGAG
CTAAAGAGGT
CCCTAGCGCT
CTTATCATGG
GGAAGCTCGG
ATCATATGCA
AGACAAAATA
AACTCGCAAC
AGCACTTGGA
GAAATGGGAC
GAATCGAGAA
AACCCTCTTT
ACGCTGGATT
ACATATCTAA
TAAAGCCGTA
AGGAGACGGG
TTCAAAAAGG
TTTAAATAAA
GGAGAAGCAA
TCAATGCATT
AGCTAGAACT
ATATTTTTTG
GACAACGTGG
AGAATTTAGA
GAACGTGCTC
TCCAAGACCA
GTTACAAAGA
GCTAGTGCAC
TAAACATAAT
TATTAACGCT
ATAAGTGTGT
GGAACACTGT
ATATATGGAA
AAAGCCGTAG
AAGAATTAAA
AGCAAGAGGA
GAATTTAGAG
AAGATTTAAT
GCCATATGCG
TGGCCGTTAG
GATGGGAACA
TATCAATTTT
CTTGGAGAAT
ACAAATTTGA
AGGATTACAT
GACACTGGGC
AAATGAATTT
ACGTCCTTTA
CGTATAAAAG
AGCCGTTTTA
TTCTTAATAT
AACGGCTCTT
TTTATAGAAA
AAATCCTTAG
CGTGGTTTTT
TTCCGAAATG
CTGGCGGTAC
CCCAAGAATT
AGAAATGAGT
AGATCAAATT
ATTCACGAAT
AGAATCAGGA
AAATCAGATC
CAACCATAAA
AACACTAGAA
CAAATTGCAA
AGTTAACTAA
CTCAACGCTA
GTAGTGGATT
TAATCCCAAA
TGAGCCAACA
GAACCAGAGC
CAGAAACAGA
ATCAGAACAA
GTAACATTGG
ATTTAGAAAT
GGAAGAAGAA
AAAAGCAATG
ACTTCGTGTG
AATAATGCAC
GAAATCGTTG
CTTATTTTTT
TTTAAAAGCG
GTATACTAGA
TATAACGAAA
CAACGAACTG
AATAGAAACG
AAAAAAGAGC
CATGACACAT
TTATAAAATG
TTTGACGACA
TTTTATAAAT
GCATAGCCCG
ATAAGATTGC
CAAACCAACG
CTTATCAGTT
AGTCAGATGA
ACTCTTCCCT
CGTAAGAAGT
TATTTAATTA
ACTTTGTTTG
AAGACGGTAT
ATAACCGTAC
TATCATTATA
TAGGGAAATC
AGAGAGTTTT
CAAGTATCTA
AGCTACTGAA
TTTAAGAATT
GTTAAGCAAT
CAATCGGAAA
TCGTTTGATT
GCTTTTTTTG
TATTCATTTA
TAGAAGGTGG
AGTTTGTATG
AATCATGATG
AATGTAAAAC
TTATATAAAA
AATAGTTTAT
TGGAGATAAG
AAAATTAGCA
AATATCTATA
CACTAGAAAC
GTTTAAGAAA
GAGTTAGAAA
AGAGAAATAT
CTACTTAGAA
ACAAAATCAG
ATAAGTATTT
TTCTTCGGAG
GGGGAAGATT
ATATATATAA
GTTAATAGAA
AATAACAAAA
TAATTTATTC
GATTAGTGGA
AAAAAATTGA
CTTATAAAGG
AAAAAAATCT
TTTTCAAAAC
ATGCAATATT
GAAACAGTTG
AATGAAAAAG
CAAACCAAGT
TAATTAAACA
ACCTATTTTA
TAGGATTTAT
AGGAAAGGAG
AACAGCTGAA
TGAATATCCC
TTTTGTTGTA
GAAACTGTGC
TTCATGACGG
CTTGTTAAAG
TACAAATTTA
AAAATAGTAA
AATTCGCTCA
ATCACTACCA
AGCCAGGTAA
AAGCAAAGGG
GCTATTTTTG
CGTATCGCTC
AAAATCAAGC
ATGATTGGCG
GTCGTGGTGT
TGTTCTGACT
TCCGAGGAAG
CGATTCAAGA
AAATCAAGAT
ACATTTACAC
ATTGGACACC
CAACGTTTAT
CGTTATGGAA
CGTATGCAGA
CGAAAACCGT
TCATACACGA
AAGGACATTC
TGAAAACAAT
TTAAGACAAA
TCAATACCTT
CTTTATTGAT
TTTGATATTC
ACACGGCAAA
AGAAACTATT
TCAGCAAGCG
ATATTTTAAC
AACCGCTATT
GATTTAGGTT
TTATGCCTAC
TATGATTATC
AAATCTGATA
AAGGTTATCA
AGCATATTTT
GTTTTAGAAA
CGCCAGTCTA
TGTGACTTCA
AAATCAGAAT
TTAAATCTGT
CAAAGCAGCC
AAAATAATTT
CGCAAAATAT
CCGAGAATAT
TTTGGAAAGT
CTTTGCCAGT
TGATCTAACG
TGTAATCATT
TTGGTATTGC
TCGCATACCA
AGAACGGACA
ATGTAGAATT
TTTTGATCCT
AATTACCGTT
ATTCTTTCAA
AGAATGGCAA
GATTGGTCTT
TCAAACAAAC
AGATAATAAG
GGCTTTACTC
GTTCAAGTCT
AACGGTTTTA
AGCGGTACAG
AAGGCAAAAA
ACAAGTAGAT
GAACCCTGGT
TTAATCTCTT
ATTGCACGAA
ACGAAATTTT
CAGGAGAAAA
GGGTTTAATA
GGGCGTAATA
ACGTCATGTT
TACCCTCTCT
TTAGCCTACT
TTAGTTCAGG
CTATTCAATC
GAAACGTGCG
AATATAATAT
GTTTGAGTTT
AATAATCGAT
TAGATCAACC
CTTAGAAGAA
AAAGAAGTAA
TCAAAATTGT
TAGAAGTGCC
TATTCAGAAA
ACTATCAAGG
GGCTAATAGG
GAATACATTA
CCATTCTTTG
CAAAGCTTGG
GTATCAAGTG
ATTTAACCAG
TAAAGATTTA
TTTGTCCGTC
AAGGGTGGTT
TAAATTCAAG
AAAAAAAGAA
GCGAACGTCA
ACGTGTTCAT
TTGTCAGAAT
GGAAAGAAGA
TTTAATGGCT
TATATTAGCG
AAAAAAGCGA
TGTATACAAG
CCTTATTTAG
TGACGACCAA
AAAAGAGATT
AGAGAAGTGC
TAGGCATTCC
TGAACGGACA
TTAGATAAAT
TGCTGAAGGT
ACTGAAGGCG
AATCAGGAAA
TTTTCTTTAA
GATTAAACCA
GGAAGAAATG
GTGGCATTCA
ACTTGCTAGT
GTTAAATCAT
TGTTGCTATC
GATCATTAAA
GTAAAAAAAG
AAGAAAAAGA
AAGCTATATA
AAGGCGCTGA
CAAATTCTTT
TGACTTAGAG
CATACATTCA
TTCAAGAGAC
TTTAAACAAG
CTAGCAGAAC
GCCCTAAAAC
GGACACACAA
CTCGATTTGT
TTAGCTATGA
TACAGGCTGA
AAATAAAACC
CGCACTATGC
CATTACATTT
ATATCTATGA
TACGTGTTTG
TTTTTTCTTT
GCTGTTTAGC
GAATGATTAG
CAGAAATATA
CAGAGTAAGA
TTTTAATTAA
TTATTAGGGG
GAGAAGGAGA
GAGTAGCCCG
AAAACTTTTA
GTTGGCTTGG
ACTGAACGAA
GTGAGGGAAA
GGCTACTAAA
ACGTCGAGGG
GCAGTGAGAG
CGAAGCGAAC
ACTTGATTTT
TTAATTTTCT
ATCTTTTATA
GGTCATTAGA
GTATACTTAT
TTGTCCTATA
AACTATTTAG
CAGCATAATA
GATTTATTGA
ATAGGTCATT
TAAGTTGAGC
ATATTAGAGG
AGGAAAATCT
TGGAGAAATA
TTTGAAGAAC
CCGATTACAT
GGATTGGATT
AGTTCTTGTG
GTTACGTGGT
TTTTAACTAA
AAGTAGTGAA
TTTTTGATTT
TTGGTGTGTG
TGTCTTGTTG
TTAGTATTTG
CTAGTCAAAG
TGATTAAATA
GAATTCCGGA
TGAGCATTCA
TCAGGCGGGC
AAGAATGTGA
ATAAAGGCCG
GATAAAACTT
GTGCTTATTT
TTCTTTACGG
TCTTTAAAAA
GGCCGTAATA
TCCAGCTGAA
CGGTCTGGTT
ATAGGTACAT
TGAGCAACTG
ACTGAAATGC
CTCAAAATGT
TCTTTACGAT
GCCATTGGGA
TATATCAACG
GTGGTATATC
CAGTGATTTT
TTTCTCCATT
TTAGCTTCCT
TAGCTCCTGA
AAATCTCGAT
AACTCAAAAA
ATACGCCCGG
TAGTGATCTT
ATTTCATTAT
GGTGAAAGTT
GGAACCTCTT
ACGTGCCGAT
CAACGTCTCA
TTTTCGCCAA
AAGTTGGCCC
AGGGCTTCCC
GGTATCAACA
GGGACACCAG
GATTTATTTA
TTCTGCGAAG
TGATCTTCCG
TCACAGGTAT
TTATTCGGCG
CAAAGTGCGT
CGGGTGATGC
TGCCAACTTA
CTGATTTAGT
GTATGATGGT
GTTTTTGAGG
TGCTCCAGTG
GCTTCTGTTT
CTATCAGCTG
TCCCTCCTGT
TCAGCTACTG
ACGGGGTGGT
GCGTAACGGC
AAAAGCACCG
CCGGACATCA
GCGCTAGCGG
AGTGTATACT
GGCTTACTAT
GTTGGCACTG
ATGAGGGTGT
CAGTGAAGTG
CTTCATGTGG
CAGGAGAAAA
AAGGCTGCAC
CGGTGCGTCA
GCAGAATATG
TGATACAGGA
TATATTCCGC
TTCCTCGCTC
ACTGACTCGC
TACGCTCGGT
CGTTCGACTG
CGGCGAGCGG
AAATGGCTTA
CGAACGGGGC
GGAGATTTCC
TGGAAGATGC
CAGGAAGATA
CTTAACAGGG
AAGTGAGAGG
GCCGCGGCAA
AGCCGTTTTT
CCATAGGCTC
CGCCCCCCTG
ACAAGCATCA
CGAAATCTGA
CGCTCAAATC
AGTGGTGGCG
AAACCCGACA
GGACTATAAA
GATACCAGGC
GTTTCCCCCT
GGCGGCTCCC
TCGTGCGCTC
TCCTGTTCCT
GCCTTTCGGT
TTACCGGTGT
CATTCCGCTG
TTATGGCCGC
GTTTGTCTCA
TTCCACGCCT
GACACTCAGT
TCCGGGTAGG
CAGTTCGCTC
CAAGCTGGAC
TGTATGCACG
AACCCCCCGT
TCAGTCCGAC
CGCTGCGCCT
TATCCGGTAA
CTATCGTCTT
GAGTCCAACC
CGGAAAGACA
TGCAAAAGCA
CCACTGGCAG
CAGCCACTGG
TAATTGATTT
AGAGGAGTTA
GTCTTGAAGT
CATGCGCCGG
TTAAGGCTAA
ACTGAAAGGA
CAAGTTTTGG
TGACTGCGCT
CCTCCAAGCC
AGTTACCTCG
GTTCAAAGAG
TTGGTAGCTC
AGAGAACCTT
CGAAAAACCG
CCCTGCAAGG
CGGTTTTTTC
GTTTTCAGAG
CAAGAGATTA
CGCGCAGACC
AAAACGATCT
CAAGAAGATC
ATCTTATTAA
TCAGATAAAA
TATTTCTAG