名称: | pUC-GW-Kan-4×SUNTag |
---|---|
别称: | |
基因长: | |
质粒宿主: | 大肠杆菌 |
质粒用途: | 基因克隆 |
片段类型: | |
片段物种: | |
原核抗性: | Kan |
筛选标记: | |
荧光标记: | |
启动子: | |
复制子: | pUC |
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。
TCGCGCGTTT
CGGTGATGAC
GGTGAAAACC
TCTGACACAT
GCAGCTCCCG
GAGACTGTCA
CAGCTTGTCT
GTAAGCGGAT
GCCGGGAGCA
GACAAGCCCG
TCAGGGCGCG
TCAGCGGGTG
TTGGCGGGTG
TCGGGGCTGG
CTTAACTATG
CGGCATCAGA
GCAGATTGTA
CTGAGAGTGC
ACCATATGCG
GTGTGAAATA
CCGCACAGAT
GCGTAAGGAG
AAAATACCGC
ATCAGGCGCC
ATTCGCCATT
CAGGCTGCGC
AACTGTTGGG
AAGGGCGATC
GGTGCGGGCC
TCTTCGCTAT
TACGCCAGCT
GGCGAAAGGG
GGATGTGCTG
CAAGGCGATT
AAGTTGGGTA
ACGCCAGGGT
TTTCCCAGTC
ACGACGTTGT
AAAACGACGG
CCAGTGAATT
GACGCGTATT
GGGATCCTGA
AGATCAAGGC
TGCTGCAGTG
AGCTGTGATC
GCACTACTGC
ACTCCAGCCT
GGGCCACAGA
GTGAACCCTG
TCTCTAAAAA
AAAATAAAAA
CAAAACAAGA
GTCCCTGGGC
CCTTTGAATA
AAAAGAGAGT
AAAGAGGCTG
GGCACAGTGG
CTCACGCCCG
TAATCCCAGC
ACTTTGGGAG
GCTGAGGCAG
GCAGATCACT
TGTGCCCAGG
AGTTTGAGGC
CAGCGTGGGC
AACATAGCAA
AATCCCATCT
CTCCAAAAAA
AAAAAAAAAA
GGAAAAGAAA
AGAAAGTGAG
TAAAGGCTTT
TGGGTTATAA
AGAGAAAGGG
AGAAGGATGA
TCCAGACTAG
AAAGAAGATT
ATTCCTTTCT
TAGCTCAACC
TTTTTGCTCC
AAAAAAAAGA
GGAGGAAGGG
GCCAAACATA
GATTATCCTG
CACCAGAGGG
CCAGCTCCTT
AAATCTGAGT
CTCATCCCAA
CCACCATCAT
TTGTAAAAAC
CAAGGGTGCA
TTCATTACCT
GCAGTGCGTG
GATCCCTGTG
GTGCCATCAT
TATCATTTGA
AGCTCCATCT
CTGAAGCCTC
TAGCTTTGTA
AGGACCTACA
GACCACTTCC
CAAGTCATTG
CAGGTTTATT
TTTGTCGCAT
TTGAGCATAA
CCTCAGATTT
GGCATACAAT
ACGAATGTCA
CCTTGAACAG
CTTCTCCTTG
AATGGGGGCC
TACTCTCCCC
AGGTGCAGTG
AGCACCGTCG
TCTTTACATG
AGTGGGCCTA
CACCACCCTG
CTGAGTAAAT
CCATCTGTTC
TGTTTCAGTA
ACTGTGGCCT
CTACAAATGG
GAACGGGACG
GAATGCTAAA
GGTATGCCAT
TTGCGTACTA
ATTTTTGACT
CCTTTTAGTG
ACCCATGCTA
ATAATGTGGA
ACCATCTCCT
ATTAAAATAT
TTTCATTTTT
CTAGGAAAAG
ACTGGTCCAA
AAATAGGAGG
AGAAACTCTG
TTACCAAGAG
GAGAAGAACA
TGCTAAATTT
CTGAACAGCC
TTAAAGGTGG
ATCTGGAGGT
GGAGGTTCTG
GAGGAGAAGA
ACTTTTGAGC
AAGAATTATC
ATCTTGAGAA
CGAAGTGGCT
CGTCTTAAGA
AAGGTTCTGG
CAGTGGAGAA
GAACTGCTTT
CAAAGAATTA
CCACCTGGAA
AATGAGGTAG
CTAGACTGAA
AAAGGGGAGC
GGAAGTGGGG
AGGAGTTGCT
GAGCAAAAAT
TATCATTTGG
AGAACGAAGT
AGCACGACTA
AAGAAAGGGT
CCGGATCGGG
TGAGGAGTTA
CTCTCGAAAA
ATTATCATCT
CGAAAACGAA
GTGGCTCGGC
TAAAAAAGGG
CAGTGGTTCT
GGATAACCCG
AACTTCAGAC
ATTTTCCCAC
AGACTTCCTG
GCCTCCTGTG
ACTCTGGAAA
GCAAAGGATT
GGCTGTGTAT
TGTCCATTGA
TTCCTGATTG
ACGCCGTCAA
AAACAAATGC
TTGTTAAGCC
CATAAGCTTT
GCCTGCTTAC
TTTCTGCCAT
TGGGTTGGTT
TGATACCACA
TTTAACATTG
ACATTTAAGT
GGAAAACCAA
GTTATCATTG
TCTTTTCTAA
GCTCAGTGTG
GATGATTGCA
TTACTTCATT
CACTGAAGTT
TTTGCCCAAA
AATTGGAAGG
TAAACAGAGA
GCTATGTTTC
TGTATCTTTT
GGTTATAGAG
TGTTCACTTC
TTTATCATAA
CAAAATTCTA
GTGTTTATAC
GAACACCCAG
AGGCAAAAGA
ATTTGGCTTA
ATTCTCACTC
CAGGTAAGTA
GCTTAACTTC
TGGGCTTCAG
TTTTCTCATC
TGTAAAATCA
GGAAGATTGG
ACTAAGTGAT
CCTGAAATGT
ATTTTTTAGC
ACTGGATTTC
TACAAATAAT
AAAACTTTCC
CATCTAGATA
ATGATGATCA
CATAGTCTTG
ATGTACGGAC
ATTAAAAGCC
AGATTTCTTC
ATTCAATTCT
GTTATCTCTG
TTTTACTCTT
TGAAATTGAT
CAAGCCACTG
AATCACTTTG
CATTTCAGTT
TATATATATA
GAGAGAAAGA
AGGTGTCTGC
TCTTACATTA
TTGTGGAGCC
CTGTGATAGA
AATATGTAAA
ATCTCATATT
ATTTTTTTTT
TAATTTTTTT
ATTTTTTATG
ACAGGGTCTC
ACTATGTCAC
CCTGGCTGGA
GTGCAGTAGT
GCGATCGCGG
CACACTGCAG
CCTTGGCTTC
CCTGGGCTCA
AGCAGTCCTC
CCACCTCAGT
CTCCCAAATA
GCTAGGACTA
CAGGCGTGCG
TGACCAAGCC
CAGCTAATTT
TTGCATTTTT
TGTAGAGATG
GGGTTTTGCC
ATGTTGCTCA
GGCTGGTCTC
AAACTCCTGA
GCACTAGCAA
TCCACCCACC
TCTATCCCAA
TGGCGCGCCG
AGCTTGGCTC
GAGCATGGTC
ATAGCTGTTT
CCTGTGTGAA
ATTGTTATCC
GCTCACAATT
CCACACAACA
TACGAGCCGG
AAGCATAAAG
TGTAAAGCCT
GGGGTGCCTA
ATGAGTGAGC
TAACTCACAT
TAATTGCGTT
GCGCTCACTG
CCCGCTTTCC
AGTCGGGAAA
CCTGTCGTGC
CAGCTGCATT
AATGAATCGG
CCAACGCGCG
GGGAGAGGCG
GTTTGCGTAT
TGGGCGCTGT
TCCGCTTCCT
CGCTCACTGA
CTCGCTGCGC
TCGGTCGTTC
GGCTGCGGCG
AGCGGTATCA
GCTCACTCAA
AGGCGGTAAT
ACGGTTATCC
ACAGAATCAG
GGGATAACGC
AGGAAAGAAC
ATGTGAGCAA
AAGGCCAGCA
AAAGGCCAGG
AACCGTAAAA
AGGCCGCGTT
GCTGGCGTTT
TTCCATAGGC
TCCGCCCCCC
TGACGAGCAT
CACAAAAATC
GACGCTCAAG
TCAGAGGTGG
CGAAACCCGA
CAGGACTATA
AAGATACCAG
GCGTTTCCCC
CTGGAAGCTC
CCTCGTGCGC
TCTCCTGTTC
CGACCCTGCC
GCTTACCGGA
TACCTGTCCG
CCTTTCTCCC
TTCGGGAAGC
GTGGCGCTTT
CTCATAGCTC
ACGCTGTAGG
TATCTCAGTT
CGGTGTAGGT
CGTTCGCTCC
AAGCTGGGCT
GTGTGCACGA
ACCCCCCGTT
CAGCCCGACC
GCTGCGCCTT
ATCCGGTAAC
TATCGTCTTG
AGTCCAACCC
GGTAAGACAC
GACTTATCGC
CACTGGCAGC
AGCCACTGGT
AACAGGATTA
GCAGAGCGAG
GTATGTAGGC
GGTGCTACAG
AGTTCTTGAA
GTGGTGGCCT
AACTACGGCT
ACACTAGAAG
AACAGTATTT
GGTATCTGCG
CTCTGCTGAA
GCCAGTTACC
TTCGGAAAAA
GAGTTGGTAG
CTCTTGATCC
GGCAAACAAA
CCACCGCTGG
TAGCGGTGGT
TTTTTTGTTT
GCAAGCAGCA
GATTACGCGC
AGAAAAAAAG
GATCTCAAGA
AGATCCTTTG
ATCTTTTCTA
CGGGGTCTGA
CGCTCAGTGG
AACGAAAACT
CACGTTAAGG
GATTTTGGTC
ATGAGATTAT
CAAAAAGGAT
CTTCACCTAG
ATCCTTTTAA
ATTAAAAATG
AAGTTTTAAA
TCAATCTAAA
GTATATATGA
GTAAACTTGG
TCTGACAGTT
AGAAAAACTC
ATCGAGCATC
AAATGAAACT
GCAATTTATT
CATATCAGGA
TTATCAATAC
CATATTTTTG
AAAAAGCCGT
TTCTGTAATG
AAGGAGAAAA
CTCACCGAGG
CAGTTCCATA
GGATGGCAAG
ATCCTGGTAT
CGGTCTGCGA
TTCCGACTCG
TCCAACATCA
ATACAACCTA
TTAATTTCCC
CTCGTCAAAA
ATAAGGTTAT
CAAGTGAGAA
ATCACCATGA
GTGACGACTG
AATCCGGTGA
GAATGGCAAA
AGTTTATGCA
TTTCTTTCCA
GACTTGTTCA
ACAGGCCAGC
CATTACGCTC
GTCATCAAAA
TCACTCGCAT
CAACCAAACC
GTTATTCATT
CGTGATTGCG
CCTGAGCGAA
ACGAAATACG
CGATCGCTGT
TAAAAGGACA
ATTACAAACA
GGAATCGAAT
GCAACCGGCG
CAGGAACACT
GCCAGCGCAT
CAACAATATT
TTCACCTGAA
TCAGGATATT
CTTCTAATAC
CTGGAATGCT
GTTTTCCCAG
GGATCGCAGT
GGTGAGTAAC
CATGCATCAT
CAGGAGTACG
GATAAAATGC
TTGATGGTCG
GAAGAGGCAT
AAATTCCGTC
AGCCAGTTTA
GTCTGACCAT
CTCATCTGTA
ACATCATTGG
CAACGCTACC
TTTGCCATGT
TTCAGAAACA
ACTCTGGCGC
ATCGGGCTTC
CCATACAATC
GATAGATTGT
CGCACCTGAT
TGCCCGACAT
TATCGCGAGC
CCATTTATAC
CCATATAAAT
CAGCATCCAT
GTTGGAATTT
AATCGCGGCC
TAGAGCAAGA
CGTTTCCCGT
TGAATATGGC
TCATACTCTT
CCTTTTTCAA
TATTATTGAA
GCATTTATCA
GGGTTATTGT
CTCATGAGCG
GATACATATT
TGAATGTATT
TAGAAAAATA
AACAAATAGG
GGTTCCGCGC
ACATTTCCCC
GAAAAGTGCC
ACCTGACGTC
TAAGAAACCA
TTATTATCAT
GACATTAACC
TATAAAAATA
GGCGTATCAC
GAGGCCCTTT
TGTC