名称: | pNZ8148-Pnis-MCS1-P32-MCS2 |
---|---|
别称: | |
基因长: | |
质粒宿主: | 乳酸菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 空载体 |
原核抗性: | Chl |
筛选标记: | Chl |
荧光标记: | |
启动子: | PnisA,P32 |
复制子: | repA |
拷贝数: | |
感受态: | MC1061 |
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。
AGATCTAGTC
TTATAACTAT
ACTGACAATA
GAAACATTAA
CAAATCTAAA
ACAGTCTTAA
TTCTATCTTG
AGAAAGTATT
GGTAATAATA
TTATTGTCGA
TAACGCGAGC
ATAATAAACG
GCTCTGATTA
AATTCTGAAG
TTTGTTAGAT
ACAATGATTT
CGTTCGAAGG
AACTACAAAA
TAAATTATAA
GGAGGCACTC
ACCATGGGTA
CTGCAGGCAT
GCGGTACCAC
TAGTTCTAGA
GAGCTCAAGC
TTTTCAAGGC
AATCTGCCTC
CTCATCCTCT
TCATCCTCTT
CGTCTTGGTA
GCTTTTTAAA
TATGGGTCGA
TCGAATTCGG
TCCTCGGGAT
ATGATAAGAT
TAATAGTTTT
AGCTATTAAT
CTTTTTTTAT
TTTTATTTAA
GAATGGCTTA
ATAAAGCGGT
TACTTTGGAT
TTTTGTGAGC
TTGGACTAGA
AAAAAACTTC
ACAAAATGCT
ATACTAGGTA
GGTAAAAAAA
TATTCGGAGG
AATTTTGAAT
CTTTGAACCA
AAGGATCCAT
TAGACCAAGG
CCTCGAGTTG
AAACGTTCAA
TTGAAATAAG
CTTGGCAATA
TTACAGCACG
TGTTGCTTTG
ATTGATAGCC
AAAAAGCAGC
AGTTGATAAA
GCAATTACTG
ATATTGCTGA
AAAATTGTAA
TTTATAAATA
AAAATCACCT
TTTAGAGGTG
GTTTTTTTAT
TTATAAATTA
TTCGTTTGAT
TTCGCTTTCG
ATAGAACAAT
CAAATCGTTT
CTGAGACGTT
TTAGCGTTTA
TTTCGTTTAG
TTATCGGCAT
AATCGTTAAA
ACAGGCGTTA
TCGTAGCGTA
AAAGCCCTTG
AGCGTAGCGT
GGCTTTGCAG
CGAAGATGTT
GTCTGTTAGA
TTATGAAAGC
CGATGACTGA
ATGAAATAAT
AAGCGCAGCG
TCCTTCTATT
TCGGTTGGAG
GAGGCTCAAG
GGAGTTTGAG
GGAATGAAAT
TCCCTCATGG
GTTTGATTTT
AAAAATTGCT
TGCAATTTTG
CCGAGCGGTA
GCGCTGGAAA
ATTTTTGAAA
AAAATTTGGA
ATTTGGAAAA
AAATGGGGGG
AAAGGAAGCG
AATTTTGCTT
CCGTACTACG
ACCCCCCATT
AAGTGCCGAG
TGCCAATTTT
TGTGCCAAAA
ACGCTCTATC
CCAACTGGCT
CAAGGGTTTG
AGGGGTTTTT
CAATCGCCAA
CGAATCGCCA
ACGTTTTCGC
CAACGTTTTT
TATAAATCTA
TATTTAAGTA
GCTTTATTTT
TGTTTTTATG
ATTACAAAGT
GATACACTAA
TTTTATAAAA
TTATTTGATT
GGAGTTTTTT
AAATGGTGAT
TTCAGAATCG
AAAAAAAGAG
TTATGATTTC
TCTGACAAAA
GAGCAAGATA
AAAAATTAAC
AGATATGGCG
AAACAAAAAG
ATTTTTCAAA
ATCTGCGGTT
GCGGCGTTAG
CTATAGAAGA
ATATGCAAGA
AAGGAATCAG
AACAAAAAAA
ATAAGCGAAA
GCTCGCGTTT
TTAGAAGGAT
ACGAGTTTTC
GCTACTTGTT
TTTGATAAGG
TAATTATATC
ATGGCTATTA
AAAATACTAA
AGCTAGAAAT
TTTGGATTTT
TATTATATCC
TGACTCAATT
CCTAATGATT
GGAAAGAAAA
ATTAGAGAGT
TTGGGCGTAT
CTATGGCTGT
CAGTCCTTTA
CACGATATGG
ACGAAAAAAA
AGATAAAGAT
ACATGGAATA
GTAGTGATGT
TATACGAAAT
GGAAAGCACT
ATAAAAAACC
ACACTATCAC
GTTATATATA
TTGCACGAAA
TCCTGTAACA
ATAGAAAGCG
TTAGGAACAA
GATTAAGCGA
AAATTGGGGA
ATAGTTCAGT
TGCTCATGTT
GAGATACTTG
ATTATATCAA
AGGTTCATAT
GAATATTTGA
CTCATGAATC
AAAGGACGCT
ATTGCTAAGA
ATAAACATAT
ATACGACAAA
AAAGATATTT
TGAACATTAA
TGATTTTGAT
ATTGACCGCT
ATATAACACT
TGATGAAAGC
CAAAAAAGAG
AATTGAAGAA
TTTACTTTTA
GATATAGTGG
ATGACTATAA
TTTGGTAAAT
ACAAAAGATT
TAATGGCTTT
TATTCGCCTT
AGGGGAGCGG
AGTTTGGAAT
TTTAAATACG
AATGATGTAA
AAGATATTGT
TTCAACAAAC
TCTAGCGCCT
TTAGATTATG
GTTTGAGGGC
AATTATCAGT
GTGGATATAG
AGCAAGTTAT
GCAAAGGTTC
TTGATGCTGA
AACGGGGGAA
ATAAAATGAC
AAACAAAGAA
AAAGAGTTAT
TTGCTGAAAA
TGAGGAATTA
AAAAAAGAAA
TTAAGGACTT
AAAAGAGCGT
ATTGAAAGAT
ACAGAGAAAT
GGAAGTTGAA
TTAAGTACAA
CAATAGATTT
ATTGAGAGGA
GGGATTATTG
AATAAATAAA
AGCCCCCCTG
ACGAAAGTCG
ACGGCAATAG
TTACCCTTAT
TATCAAGATA
AGAAAGAAAA
GGATTTTTCG
CTACGCTCAA
ATCCTTTAAA
AAAACACAAA
AGACCACATT
TTTTAATGTG
GTCTTTTATT
CTTCAACTAA
AGCACCCATT
AGTTCAACAA
ACGAAAATTG
GATAAAGTGG
GATATTTTTA
AAATATATAT
TTATGTTACA
GTAATATTGA
CTTTTAAAAA
AGGATTGATT
CTAATGAAGA
AAGCAGACAA
GTAAGCCTCC
TAAATTCACT
TTAGATAAAA
ATTTAGGAGG
CATATCAAAT
GAACTTTAAT
AAAATTGATT
TAGACAATTG
GAAGAGAAAA
GAGATATTTA
ATCATTATTT
GAACCAACAA
ACGACTTTTA
GTATAACCAC
AGAAATTGAT
ATTAGTGTTT
TATACCGAAA
CATAAAACAA
GAAGGATATA
AATTTTACCC
TGCATTTATT
TTCTTAGTGA
CAAGGGTGAT
AAACTCAAAT
ACAGCTTTTA
GAACTGGTTA
CAATAGCGAC
GGAGAGTTAG
GTTATTGGGA
TAAGTTAGAG
CCACTTTATA
CAATTTTTGA
TGGTGTATCT
AAAACATTCT
CTGGTATTTG
GACTCCTGTA
AAGAATGACT
TCAAAGAGTT
TTATGATTTA
TACCTTTCTG
ATGTAGAGAA
ATATAATGGT
TCGGGGAAAT
TGTTTCCCAA
AACACCTATA
CCTGAAAATG
CTTTTTCTCT
TTCTATTATT
CCTTGGACTT
CATTTACTGG
GTTTAACTTA
AATATCAATA
ATAATAGTAA
TTACCTTCTA
CCCATTATTA
CAGCAGGAAA
ATTCATTAAT
AAAGGTAATT
CAATATATTT
ACCGCTATCT
TTACAGGTAC
ATCATTCTGT
TTGTGATGGT
TATCATGCTG
GATTGTTTAT
GAACTCTATT
CAGGAATTGT
CAGATAGGCC
TAATGACTGG
CTTTTATAAT
ATGAGATAAT
GCCGACTGTA
CTTTTTACAG
TCGGTTTTCT
AATGTCACTA
ACCTGCCCCG
TTAGTTGAAG
AAGGTTTTTA
TATTACAGCT
CC