名称: | pCR-BluntII-TOPO-FRMPD1(1点突变1同义突变)(human) |
---|---|
别称: | NM_014907.3(1点突变1同义突变);FRMD2 |
基因长: | 4320bp |
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。
ATAGGTGACC
TATAGATACT
CCAAGCTATG
CATCAAGCTT
GGTACCGAGC
TCGGATCCAC
TAGTAACGGC
CGCCAGTGTG
CTGGAATTCG
CCCTTGGCGG
CACCTCCTCT
GCAGGTAAAT
GGAAGAGCTG
GAGACCAGTT
TATTCCAGAC
ACGGAAAGCA
CATAGAATAG
AACAAATGGT
GGCAAGATGG
CTTCGGCGCT
CCCGGGACAG
CTCGGCCCGG
GCTAAAGTTG
CTGCAGCTGA
TGGGCCCGCC
AGGAACCCAA
CTCAGACCCT
CATCCCTGTG
CGACACACAG
TAAAGATAGA
CAAAGACACC
CTCCTCCAGG
ACTATGGATT
TCACATTTCT
GAGAGCCTTC
CCCTTACAGT
GGTGGCTGTC
ACAGCAGGAG
GCTCTGCTCA
CGGCAAGCTT
TTCCCTGGTG
ATCAGATCCT
CCAAATGAAC
AATGAGCCTG
CTGAAGACCT
TTCCTGGGAA
CGAGCAGTCG
ATATTCTCAG
GGAAGCAGAA
GATTCTCTTT
CAATCACAGT
TGTTCGCTGC
ACGTCGGGAG
TCCCTAAATC
GTCCTTCCTG
ACCGAGGAGA
AGAGAGCCAG
ACTGAAGACC
AACCCCGTGA
AGGTGCACTT
TGCTGAAGAA
GTGCTCATCA
GTGGACACAG
CCAGGGTAAT
TCTCTGCTGT
GTATGCCCAA
CGTGCTCAAG
CTATACTTGG
AGAATGGACA
GACCAAAGCT
TTCAAGTTTG
AGGCAAACAC
AACCGTGAAG
GACATCATCC
TCACCGTGAA
GGAGAAGCTG
TCCATCCGAA
GTATCGAGTA
CTTTGCACTG
GCCCTGGAAG
AGCAGTACAG
CATCTCCCGG
CTGCACCTGC
TGCACGAAGA
GGAACTCATC
CAGCAGGTGG
TAGAAAGGGA
GGAGTCACAT
GACTACCGCT
GCCTCTTCAG
GGTCTGTTTT
GTTCCCAAGG
ACCCCCTGGA
CCTCCTGAAA
GAAGACCCCG
TGGCCTTTGA
ATACCTCTAT
CTGCAGAGCT
GCAGCGATGT
GCTCCAGGAG
CGCTTTGCTG
TAGAAATGAA
ATGTAGCTCT
GCACTCCGAC
TCGCGGCTCT
GCACATCCAG
GAACGGATCT
ACGCCTGTGC
CCAGCCACAG
AAGATCTCTT
TAAAGTATAT
AGAGAAAGAC
TGGGGAATAG
AGAACTTTAT
ATCTCCAACT
TTACTCCGAA
ACATGAAAGG
CAAAGACATC
AAGAAAGCCA
TTAGCTTCCA
CATGAAGAGG
AACCAGAATT
TGCTGGAACC
CCGACAGAAG
CAACTTATTT
CTGCTGCCCA
GCTACGTTTA
AATTATCTAC
AGATCCTCGG
AGAACTCAAG
ACATATGGTG
GAAGAATCTT
TAATGCTACT
TTAATGTTAC
AGGATAGAGA
ATCCTACATT
GCCCTTCTAG
TTGGAGCCAA
GTATGGGATT
AGCCAGGTTA
TCAATAGCAA
ACTCAACATC
ATGTCCACAT
TGGCAGAGTT
TGCAAACATC
AGCCGTGTAG
AGCTAACGGA
AGAGTCTGAG
AAAGTGAGCG
TCGTCAAAGT
GTATCTTCAG
GACGTCAAGG
TTCTGACTTT
GCTGCTGGAA
TCCAACAGTG
CAAAAGACCT
AGCCTGCCTG
ATTGCTGGGT
ACTACAGGCT
GTTGGTTGAC
CCAGTTACCT
CCATTTTCCT
CTGGCCTGGA
AACAAACAAC
AAGCGCACCG
GGTATCTGCA
GAAGAAGGCT
ATGAATCCAG
GGCCTGCAGT
GACTCAGAGG
AGTCCTCTGA
GGTGGACTGC
GTACTCGAAC
CTCTCTCTGA
CAGGCGCCTG
GTGAAACTGG
CACCCTGCAG
ATCACTCATA
AAGGAGGAGC
AGCCTCCTGG
GAACAGCCCC
ACACCTGAGG
TGGCTAGGAG
GGGCCCCAGC
ACCTGCGGGG
CCAGCAGCAC
GACAGACAGT
GCCGAGTCCG
AGGCGTCCGA
CTCAGCCAAC
ACTGAGAGCC
GCGGCTACAG
GACCAGTGGC
TCGAGTGAGT
CCATGGACGC
TCTGGAAGAG
GATGACTTAG
ACACCTGCTC
CTCCAGCAGG
TCCACCTTCT
TCCACTTTGG
CTCGCCAGGC
CTCGCAGAGA
GCATTGACTC
TGACAGCCAG
GAGGAGAGAA
GCGGGATTGA
AACCAGTGGC
TTCCTCTGTC
TCCTGGACCT
GGCCCAGAGA
GCCAACCCCC
AGTGCCAGAA
GACAGAGTTT
TCCGAGAGTG
CTGCTTTGGA
GACATTTGGC
TGGGCACCAG
AACTGAGCAC
AGTCAGGCTG
GACCCCAGGC
TGTATGAAGG
CAGCCACGCC
GACTACTACA
GCCTGTGTTC
CAGTGTCTCC
CCGGCCAGCT
ACCTGAGTGA
CAGTTCCGAG
AGTACAGCTT
CCCGGCAAGG
GGGAGCCCCG
CCAGCCTGGG
GTCAGCAGGG
CTGGACTGAG
GCCCAGCCCA
GTTCCATGCT
GGAACCCCTG
GCCCTGCACC
CACCACTGGC
CTTTGAGGAT
GGCAGCTCAG
ATGAGGAATA
CTATGACGCG
GCTGATAAGC
TCACTCCCCC
AGGCCCCCCG
TCAGGGCCCA
GAGATGTTTC
TACTGCAGAA
CCCAGTGCCA
CAAGCTTGCA
GAATAAGGCC
AGCACTTCTA
GCCCTGAGAA
CAGCCTGCCT
TGTGGGCCAG
ATGGAAGACA
GCCAAGCAGG
AGGGGAGGGG
TGAAGAAATA
TGCCAAGACC
CTGAGGAAAA
GAAGGTCTTT
CCTACAGACC
GACGACACCT
CTCAGGTCTC
ATTTCCCCTG
GTGCCATCAG
CCTCCCTGGA
GAGTGTAGAC
GACGTGTGCT
ACTATGACAG
GGAGCCCTAC
CTGGCCCTTG
GTGCACCCTC
CCCAACTGTG
TCCTCTCTGC
AGGACATGCA
GGGTGAGCCT
GGCCTTCTGG
AGACCAAGGC
CTTGGGGCTG
CTGGCTCCTC
TGAGGGAGAC
CAAGAGCACA
AACCCAGCCT
CCAGGGTCAT
GGAGATGGAG
CCCGAGACCA
TGGAAACCAA
ATCAGTCATC
GACTCTCGAG
TGTCTTCTAT
TTCTGCCATT
CGCTTCCGGA
TTGACCCCAA
CAATAAAGAG
AATTCTGGTG
TTGTCCCTGC
TGCCAGCTCC
TCAGCAAGCA
CTCCTCACTG
TTCTAACCCA
GGTTCATCTG
GCCCAGATAC
TGCTCAGGCA
AGGCCTTCCC
AAATCTTACC
TCTATCTCAA
GACCTGGATG
GGATTGCCCC
CAAAGAACCA
ACCATAGAGC
ATGGAGACAG
CTCCTTCTCC
CTCTCCAGTG
GTGACCCTAA
CCCAGACAGA
GCCTGCCTGG
CCAGCAACCC
AGGACTAAAT
AATGTCTCTC
AAGGAGACAC
ACTAGAGCTC
CAGTTGGAGC
CCCATGTCCA
GTTGGAAATG
GGATTGGAAT
CTTTTTGTAC
AAATCATATA
CAAGAAACTG
CCCCCAAATA
CACAGAGCCT
TTGTTGTCTC
CTAGAGATGA
GCCTAGAAGT
GATGAATGTG
GAATAAATCC
AGGAGAGAAG
ATAGCTTCTA
TCCCTACAAA
GGAAGAGCCA
CAAGGACAAC
TATCTCTGGA
AAGAGACAGA
GAAGTTACAA
ACAAAAATGG
CACCAACGTA
TTTCAGGAGG
AATCTAGGAA
GGATTCAGGT
GACTCCCCGG
GTGATGTGTC
AAATAATGTT
TCACAGACTC
TTGATATTAG
CTCTCCAGCT
GGTAAAATAG
TAACCTCCCT
TTCTTTAGAT
GCTCCTGTAA
CAGGGACCGA
GCAGATCCCA
CCACATCCCC
CTAGAGACCC
TCAAGGACAG
AGCAGAGAAC
CCCCAGGGCA
AGGCTGCCAG
GCTCAAGAAC
AAAAACTATT
CGTAGAGTTG
GATTTAGACC
CTGATTTCTT
TCTTGGGAAG
CAGACAGTTT
CACCAGCCGT
CCCTCCAGAG
GGGATCAAGG
CAGAGGCACC
TAACCATGTG
ACAGGGCAAG
ATATAGCCCC
TAGGGACAGC
CCTGAGTGGG
TCTGTTTTAA
TCCTGAGCCT
TCCCTGCCAG
AACCACTACC
ATGTCCACAA
GAGGATCCTC
ACTTAGAAAC
TTCAAACCAT
TGCTTACTCT
CAGAAGGCAA
AAGTGACAGC
TCTAGCATCT
GCCTTTCTGC
TGAGAAGTCT
TTTCTGTGCT
TTGCCCCAGA
AAGCCATCCT
GAAGTCTCTG
CCAGTCTCAG
GGTGGCCACA
TCTTTGGGTT
TTGCAGGCAT
GAATGAGATG
GTGGCTCCCA
GGATAGGGAT
GGACCAGTGC
AGCTGTCAGT
TCTCCTATGC
CACATGCTTC
CGTGGCCCGC
AGCCTGAGAC
AGAGGAAGAA
GACAGGGACT
TGGAAGCACA
CCCCATGGCC
CCCCTCACCT
CACCGCCCTC
TGCGGGAAGC
CCGGTGGTTC
TGCCCTGGAG
GCCTGCCCGA
GCCCACAGCT
GCACCACCGC
ACCCCTGTCG
AGGAAAAGCC
ACATCTGGCC
AGAGTACTGC
TCCAGGGCAC
TGAGACAGCT
GAAAGCCACC
CCTGCCAGCA
CCCCTGAGGG
CTTCATCCAA
CTCATGGAGA
GCTTGCTGGA
GCTACAAGAC
ATTTTAGAAA
CTTCCTGGGG
GGTTGGAAAC
AAACATCCCC
CAGAGAAGTG
CACCTGGCAC
TTTACCGAAA
GCCGGAGCCG
CCTCTGCATG
GGCTCCCAGA
AGCTCCTGTC
GAGCTGTCGG
CATGTGATCA
GAATGGACCA
GTCCCCCGAA
GAGATGCAGG
GGGCCGTGCG
TGACACCTTC
CAGCACCTGG
TCCAGCTGGC
CGGCCTGTGC
TTTCAGTTCA
CAGACTGTAG
CCGCTGCTCC
GCCCGGCACA
GGGAGGCAGC
GGGGAACCTG
AGGGATGTGG
TGTACACCTA
CCATCAGTTT
ATAGAGGCTG
CTAAATCGAC
CTGCGAGAGA
GGCTACCACG
ACCTGAGTGT
GAAACTCCTG
GCCCGTCAGT
GCACGGCCCT
CACGGCCGCC
GTGTTCTGTT
TGACCCAGAA
GTTCCGGGCA
TCCACGGCCC
TGTAAACAGG
TCAACGGCCC
AAGGGCCTCC
TGCCCTGTCC
TGCCTTGGAC
ACTTCCCTGA
GAAGCCCCTT
CCACTCTCCC
ACCCACCCTC
AATGGACGAA
GGGCGAATTC
TGCAGATATC
CATCACACTG
GCGGCCGCTC
GAGCATGCAT
CTAGAGGCCC
AATT