名称: | pCR-BluntII-TOPO-UBE4A(human) |
---|---|
别称: | NM_001204077.2;BC112367.1;E4; UBOX2; UFD2 |
基因长: | 3201bp |
质粒宿主: | 大肠杆菌 |
质粒用途: | 基因模板 |
片段类型: | cDNA |
片段物种: | 人 |
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: | M13F:TGTAAAACGACGGCCAGT |
反向引物: | M13R:CAGGAAACAGCTATGACC |
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。
TGCTCGAGCG
GCCGCCAGTG
TGATGGATAT
CTGCAGAATT
CGCCCTTTGA
GTCGGAAGTG
GGAACCCTTC
GGCCGCTGAG
ATTCTGTCGT
GTCGTCGCTG
CTGGCACTTC
AGGCTCTGCC
TCTCCCACTA
GGTCTGGATG
GAGGATACCT
TAAAGTGAAA
TGACAGACCA
GGAGAATAAC
AACAACATCT
CAAGTAACCC
CTTTGCTGCT
CTTTTTGGCT
CCCTGGCTGA
TGCCAAACAG
TTTGCGGCAA
TCCAAAAAGA
GCAGCTGAAG
CAACAATCTG
ATGAACTCCC
AGCTAGCCCA
GATGACTCGG
ATAATAGCGT
GTCAGAGAGC
CTGGATGAAT
TCGATTACTC
TGTGGCTGAG
ATTAGCCGCT
CATTCCGATC
ACAGCAGGAA
ATATGTGAGC
AACTCAACAT
CAATCACATG
ATCCAAAGGA
TCTTCCTTAT
TACTCTGGAC
AACAGTGATC
CCAGCTTGAA
AAGCGGGAAT
GGCATCCCTA
GCCGTTGTGT
GTATTTGGAA
GAAATGGCAG
TAGAGCTAGA
AGATCAAGAC
TGGCTTGATA
TGAGCAATGT
TGAGCAGGCC
CTCTTCGCTC
GCTTATTACT
TCAAGATCCA
GGCAACCACT
TAATTAACAT
GACTTCTTCT
ACAACGCTAA
ATCTCTCTGC
TGATCGAGAT
GCAGGAGAGA
GGCACATTTT
TTGTTACCTT
TACTCCTGCT
TCCAGAGAGC
CAAGGAAGAG
ATTACCAAAG
TTCCAGAGAA
CCTGCTACCC
TTTGCAGTGC
AGTGCAGAAA
CCTCACTGTG
TCCAATACCC
GAACAGTTCT
TCTCACCCCA
GAGATCTATG
TTGACCAAAA
CATCCATGAG
CAACTGGTAG
ATTTGATGTT
AGAAGCCATC
CAGGGAGCCC
ATTTTGAAGA
TGTAACTGAG
TTTCTGGAAG
AGGTCATTGA
AGCCTTGATA
TTGGATGAGG
AAGTTAGAAC
ATTTCCAGAA
GTCATGATTC
CAGTGTTTGA
TATTTTATTG
GGCCGAATAA
AAGATCTAGA
GCTCTGTCAG
ATCCTTTTGT
ATGCATATCT
GGATATTCTT
CTCTATTTCA
CTAGGCAAAA
AGATATGGCA
AAGGTTTTTG
TAGAATACAT
TCAGCCCAAG
GACCCTACCA
ATGGGCAAAT
GTACCAGAAG
ACCTTGCTGG
GAGTAATTCT
GAGTATCTCC
TGCTTATTAA
AGACTCCGGG
TGTTGTAGAA
AATCATGGCT
ACTTTTTGAA
TCCATCTCGT
TCCAGCCCCC
AGGAGATCAA
AGTACAGGAG
GCCAACATCC
ATCAGTTCAT
GGCTCAGTTC
CACGAAAAGA
TCTACCAGAT
GCTGAAGAAC
TTACTCCAGC
TCTCTCCAGA
AACCAAACAC
TGTATCTTGT
CCTGGCTTGG
AAACTGTTTG
CATGCAAATG
CAGGCCGCAC
CAAGATTTGG
GCCAATCAGA
TGCCAGAAAT
CTTTTTCCAA
ATGTATGCCT
CAGATGCTTT
CTTTCTGAAT
CTGGGTGCTG
CTCTCCTGAA
GCTATGCCAG
CCATTTTGCA
AACCCAGATC
CTCTCGGCTC
CTCACCTTTA
ATCCCACATA
CTGTGCCCTC
AAGGAGTTGA
ATGATGAAGA
ACGAAAAATT
AAAAATGTAC
ACATGAGAGG
TTTGGACAAA
GAAACCTGTT
TGATCCCAGC
TGTGCAGGAG
CCGAAGTTTC
CACAGAACTA
CAACCTTGTA
ACAGAGAACC
TTGCTCTGAC
AGAGTACACC
TTGTACTTGG
GATTTCACAG
GTTGCATGAT
CAGATGGTAA
AAATCAACCA
AAATCTGCAT
CGGCTGCAGG
TTGCCTGGCG
GGATGCTCAG
CAAAGTTCTA
GCCCTGCTGC
TGACAATCTT
CGTGAGCAGT
TTGAACGACT
GATGACCATC
TATCTTTCTA
CCAAGACTGC
CATGACAGAG
CCACAAATGC
TACAAAACTG
CCTAAACTTG
CAGGTGTCCA
TGGCTGTTCT
ACTGGTTCAA
CTGGCCATAG
GCAATGAGGG
CTCACAGCCA
ATAGAGCTAA
CCTTTCCTTT
GCCAGATGGC
TACAGCTCTT
TGGCTTATGT
GCCAGAATTT
TTTGCAGATA
ACCTGGGTGA
TTTTCTCATT
TTTCTCCGCC
GCTTTGCCGA
TGACATTTTG
GAGACATCAG
CAGATTCCCT
GGAGCATGTC
CTTCACTTTA
TCACCATTTT
CACTGGAAGC
ATAGAAAGAA
TGAAGAATCC
CCACCTGAGG
GCCAAACTAG
CAGAGGTGTT
GGAAGCAGTG
ATGCCCCACC
TGGATCAGAC
CCCAAATCCC
TTGGTATCCA
GTGTGTTCCA
CCGGAAACGT
GTGTTCTGCA
ACTTTCAGTA
TGCACCCCAA
CTTGCAGAGG
CTCTAATCAA
GGTTTTTGTG
GACATCGAAT
TTACAGGAGA
CCCCCATCAA
TTTGAACAGA
AGTTTAATTA
CCGCCGTCCC
ATGTATCCTA
TCCTAAGATA
CATGTGGGGG
ACAGATACCT
ATCGGGAGAG
CATTAAGGAT
TTGGCTGACT
ATGCCTCTAA
GAATTTAGAA
GCCATGAATC
CCCCACTTTT
CCTCCGCTTT
CTTAACCTGC
TAATGAATGA
TGCCATCTTC
CTTTTGGATG
AAGCCATACA
GTATTTGAGC
AAGATAAAGA
TTCAGCAAAT
TGAGAAGGAT
CGAGGTGAAT
GGGATAGTCT
GACTCCAGAA
GCCCGCCGAG
AAAAGGAGGC
TGGCCTACAG
ATGTTTGGAC
AGCTGGCACG
TTTCCATAAC
ATCATGTCCA
ATGAAACAAT
CGGTACCCTT
GCCTTTCTCA
CATCAGAGAT
CAAGTCACTC
TTTGTGCATC
CCTTCCTGGC
TGAGCGCATC
ATCTCCATGT
TGAACTACTT
CCTGCAACAC
CTGGTTGGCC
CCAAGATGGG
TGCCTTAAAA
GTCAAGGACT
TCAGCGAATT
TGACTTCAAA
CCCCAGCAGC
TTGTATCAGA
TATCTGCACT
ATCTACTTAA
ATCTTGGGGA
TGAGGAGAAT
TTCTGTGCCA
CTGTGCCCAA
GGATGGACGT
TCCTATTCCC
CAACTCTCTT
TGCACAGACA
GTTCGAGTCT
TGAAGAAAAT
AAATAAGCCT
GGGAATATGA
TTATGGCTTT
CAGCAACTTG
GCAGAGAGAA
TCAAGTCTCT
TGCAGACCTC
CAACAACAGG
AAGAGGAAAC
CTATGCAGAT
GCCTGTGATG
AGTTCCTGGA
TCCCATTATG
AGCACACTGA
TGTGTGACCC
TGTGGTGCTG
CCATCTTCCA
GAGTCACTGT
GGATAGATCC
ACCATTGCAA
GACATTTGCT
CAGTGACCAA
ACAGATCCCT
TTAACCGTAG
TCCCCTCACC
ATGGACCAGA
TCCGGCCAAA
CACAGAACTA
AAAGAAAAAA
TCCAACGGTG
GCTTGCAGAG
AGGAAACAAC
AAAAGGAGCA
ACTTGAATAG
ATACTGTGAA
CTAACCAAAC
CAAAACCAAC
CCCAGAGTGC
AGATAAACAA
TTGTTTGTGG
TTTCTCTCTT
TCTGGTTCTG
TTCCTTTTCT
TTCTTCTTTT
CTTTTTCTTT
TTTTTTTTTT
TTTTACTAAA
TTAGAGAACT
GCTCTTGCTG
AAATTATGAT
AGTTAAGATT
CCTAAGAACT
TGACAATGCT
CCCCTGGCTT
GCAGGAAATT
ATACTTATTA
TAGCATCACC
AAGTTTAGAA
ATCATCACTA
ATTTTCACCC
TCTGTTGTCT
CTTCATATTC
TTCTTCCTTT
TCCTAAATGA
AATTATATTA
TTTCAACTAC
TAAAACTTCC
CGCCACCCTG
CTATTTCTCT
TCCAATTACT
GTTCAAACAT
TTTTGGTGAG
TGCTGCTTTT
ATAAATATGT
GATGTCACAT
ATTTCAGTGA
CAGCTGATGT
TATCAGAAAA
ATCCTGTTAT
CCTGTGTATT
TACTAGTCCT
CATGATCTAG
ACTTAACCCC
TTTTCTGTGT
TACAGAGTAA
CTTCACATAA
AAGCAGATAC
CTTTAAGTTT
GTGTAGACTT
CTGAATAAGA
TGATAGACGA
GAATTTTTTT
TAAAAAAGGA
ATTTAAAGGA
ATCTTGTGCA
ACTTCTGTTG
ATAGCTCTTA
ATTTTGTTAT
ACAGTCTTTT
TAATGAGGAT
TGGTAGGGCA
ATCCTACCAA
TTGATGGGGG
TGAGGAGACT
GTTGGGCCCT
TATTCTCTTA
TGAAAATCTG
TTTCTACAAG
GACTAGGCCT
AAGTAGATTT
AGGCAACGGG
TATCATATCC
ATGAAAAATG
TCATTTTAAG
ACACTAATAT
CAACAGTATA
TCTCAGTGTG
TGAGATATAT
AAATACACAT
ACACCCCATG
TGCTTTTTTG
TTGGTTTAGT
GTAAACTGAG
TACCACTTTA
TATTTTCTCT
TCCATAGTGA
GATGTATGCA
GTATGTGGCC
ATGTTTACTA
ACATACTCAT
TCTTCACAAA
ATTCTGAGAT
TATAAAATGT
ATATAGTTAA
TATTTGTTTG
AGCTTGTGAC
CTGACTTCTA
AGAGCTACTT
CTAACACAGC
CTTAGGTCTT
CAATTAACAA
AGTAGGAAGT
TGAGCACAAC
CTTGTCCACA
GCCATTTTAA
ATATCCACTT
AGCCCTGTGT
AGTTGGTAGG
TGGACACCAC
TTTCATAAGT
GAACTTGAGC
TCACTACTCA
ATTGTGCAGC
TTAATATGCT
GACTAGGGAC
ATTCCCCTAT
GGATTATCTT
TATATCACTG
TCTTTCTAAG
CCCAGAGATG
TCATAAGTGA
CAAGAAAAGT
GATAGGATCA
AGAAATGGGT
GTCACTGCTT
CATTTGGAAC
ATGTTATTAA
CTATGGTCTC
TTTCAAATAA
ATAGTAGTTT
TATATTTCAA
CACAAGTTGC
TATAAGCAGT
CCTTGATGGG
TTTTTGATTG
CATCAGCTGG
AAAGCCTCAA
ATCTAAGAGG
GCTTCCCATC
CTAGATATAA
AATAGGTGTT
GCCTATTGCT
GTGCTTATAA
AATGAAAAAG
GAAATTGAGG
ACACTTTTGC
AAATGCCAGA
ATGTAAGATT
CATTCAGTGT
GCTCCCTGGG
CCTTTATGGC
ATGGGTTGAC
AGGATTTGTT
TATTTTCTAA
AATTAGCTTC
ATTCAATATT
TATCATCCTC
CTTTCCCTCT
CTGAGAATGA
ACTATGTATA
AAATAAGCTT
CTGCCTATTT
GCATTTATCT
TCCAAACCCA
ATCTAGTAGG
ATGTTCTCAT
TTTAAAAACG
AGGGGAAAAG
ACCAGAGTTT
TTCAGGAGAA
AACTGGAGGA
AAATGGGCAC
AAAAACTCAG
AAGGCAGCTA
TTCCCAGCAG
CTTCCTAGTT
AACAACCCCC
ATGCTGCCTC
CAGTCTTTTG
TGAAAGGGCG
AATTCCAGCA
CACTGGCGGC
CGTTACTAGT
GGATCCGAGC
TCGGTACCAA
GCTTGATGCA
TAGCTTGAGT
ATCT