名称: | pLVshRNA-U6-CCNDBP1(human)-shRNA1-PGK-EGFP(2A)Puro |
---|---|
别称: | |
基因长: | |
质粒宿主: | 哺乳细胞,慢病毒 |
质粒用途: | 干扰沉默 |
片段类型: | shRNA |
片段物种: | 人 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | 绿色 |
启动子: | U6 |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 37度 |
背景载体: | |
正向引物: | U6-F(发夹结构):GAGGGCCTATTTCCCATGA |
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。
TGGAAGGGCT
AATTCACTCC
CAAAGAAGAC
AAGATATCCT
TGATCTGTGG
ATCTACCACA
CACAAGGCTA
CTTCCCTGAT
TAGCAGAACT
ACACACCAGG
GCCAGGGGTC
AGATATCCAC
TGACCTTTGG
ATGGTGCTAC
AAGCTAGTAC
CAGTTGAGCC
AGATAAGGTA
GAAGAGGCCA
ATAAAGGAGA
GAACACCAGC
TTGTTACACC
CTGTGAGCCT
GCATGGGATG
GATGACCCGG
AGAGAGAAGT
GTTAGAGTGG
AGGTTTGACA
GCCGCCTAGC
ATTTCATCAC
GTGGCCCGAG
AGCTGCATCC
GGAGTACTTC
AAGAACTGCT
GATATCGAGC
TTGCTACAAG
GGACTTTCCG
CTGGGGACTT
TCCAGGGAGG
CGTGGCCTGG
GCGGGACTGG
GGAGTGGCGA
GCCCTCAGAT
CCTGCATATA
AGCAGCTGCT
TTTTGCCTGT
ACTGGGTCTC
TCTGGTTAGA
CCAGATCTGA
GCCTGGGAGC
TCTCTGGCTA
ACTAGGGAAC
CCACTGCTTA
AGCCTCAATA
AAGCTTGCCT
TGAGTGCTTC
AAGTAGTGTG
TGCCCGTCTG
TTGTGTGACT
CTGGTAACTA
GAGATCCCTC
AGACCCTTTT
AGTCAGTGTG
GAAAATCTCT
AGCAGTGGCG
CCCGAACAGG
GACTTGAAAG
CGAAAGGGAA
ACCAGAGGAG
CTCTCTCGAC
GCAGGACTCG
GCTTGCTGAA
GCGCGCACGG
CAAGAGGCGA
GGGGCGGCGA
CTGGTGAGTA
CGCCAAAAAT
TTTGACTAGC
GGAGGCTAGA
AGGAGAGAGA
TGGGTGCGAG
AGCGTCAGTA
TTAAGCGGGG
GAGAATTAGA
TCGCGATGGG
AAAAAATTCG
GTTAAGGCCA
GGGGGAAAGA
AAAAATATAA
ATTAAAACAT
ATAGTATGGG
CAAGCAGGGA
GCTAGAACGA
TTCGCAGTTA
ATCCTGGCCT
GTTAGAAACA
TCAGAAGGCT
GTAGACAAAT
ACTGGGACAG
CTACAACCAT
CCCTTCAGAC
AGGATCAGAA
GAACTTAGAT
CATTATATAA
TACAGTAGCA
ACCCTCTATT
GTGTGCATCA
AAGGATAGAG
ATAAAAGACA
CCAAGGAAGC
TTTAGACAAG
ATAGAGGAAG
AGCAAAACAA
AAGTAAGACC
ACCGCACAGC
AAGCGGCCGG
CCGCTGATCT
TCAGACCTGG
AGGAGGAGAT
ATGAGGGACA
ATTGGAGAAG
TGAATTATAT
AAATATAAAG
TAGTAAAAAT
TGAACCATTA
GGAGTAGCAC
CCACCAAGGC
AAAGAGAAGA
GTGGTGCAGA
GAGAAAAAAG
AGCAGTGGGA
ATAGGAGCTT
TGTTCCTTGG
GTTCTTGGGA
GCAGCAGGAA
GCACTATGGG
CGCAGCGTCA
ATGACGCTGA
CGGTACAGGC
CAGACAATTA
TTGTCTGGTA
TAGTGCAGCA
GCAGAACAAT
TTGCTGAGGG
CTATTGAGGC
GCAACAGCAT
CTGTTGCAAC
TCACAGTCTG
GGGCATCAAG
CAGCTCCAGG
CAAGAATCCT
GGCTGTGGAA
AGATACCTAA
AGGATCAACA
GCTCCTGGGG
ATTTGGGGTT
GCTCTGGAAA
ACTCATTTGC
ACCACTGCTG
TGCCTTGGAA
TGCTAGTTGG
AGTAATAAAT
CTCTGGAACA
GATTTGGAAT
CACACGACCT
GGATGGAGTG
GGACAGAGAA
ATTAACAATT
ACACAAGCTT
AATACACTCC
TTAATTGAAG
AATCGCAAAA
CCAGCAAGAA
AAGAATGAAC
AAGAATTATT
GGAATTAGAT
AAATGGGCAA
GTTTGTGGAA
TTGGTTTAAC
ATAACAAATT
GGCTGTGGTA
TATAAAATTA
TTCATAATGA
TAGTAGGAGG
CTTGGTAGGT
TTAAGAATAG
TTTTTGCTGT
ACTTTCTATA
GTGAATAGAG
TTAGGCAGGG
ATATTCACCA
TTATCGTTTC
AGACCCACCT
CCCAACCCCG
AGGGGACCCG
ACAGGCCCGA
AGGAATAGAA
GAAGAAGGTG
GAGAGAGAGA
CAGAGACAGA
TCCATTCGAT
TAGTGAACGG
ATCTCGACGG
TATCGCCTTT
AAAAGAAAAG
GGGGGATTGG
GGGGTACAGT
GCAGGGGAAA
GAATAGTAGA
CATAATAGCA
ACAGACATAC
AAACTAAAGA
ATTACAAAAA
CAAATTACAA
AAATTCAAAA
TTTTCGGGTT
TATTACAGGG
ACAGCAGAGA
TCCAGTTTAT
CGATCTGGGC
AGGAAGAGGG
CCTATTTCCC
ATGATTCCTT
CATATTTGCA
TATACGATAC
AAGGCTGTTA
GAGAGATAAT
TAGAATTAAT
TTGACTGTAA
ACACAAAGAT
ATTAGTACAA
AATACGTGAC
GTAGAAAGTA
ATAATTTCTT
GGGTAGTTTG
CAGTTTTAAA
ATTATGTTTT
AAAATGGACT
ATCATATGCT
TACCGTAACT
TGAAAGTATT
TCGATTTCTT
GGCTTTATAT
ATCTTGTGGA
AAGGACGAGG
ATCCGGCTCA
TAATCCCATG
CCTTGCTTCA
AGAGAGCAAG
GCATGGGATT
ATGAGCTTTT
TGAATTCTAC
CGGGTAGGGG
AGGCGCTTTT
CCCAAGGCAG
TCTGGAGCAT
GCGCTTTAGC
AGCCCCGCTG
GGCACTTGGC
GCTACACAAG
TGGCCTCTGG
CCTCGCACAC
ATTCCACATC
CACCGGTAGG
CGCCAACCGG
CTCCGTTCTT
TGGTGGCCCC
TTCGCGCCAC
CTTCTACTCC
TCCCCTAGTC
AGGAAGTTCC
CCCCCGCCCC
GCAGCTCGCG
TCGTGCAGGA
CGTGACAAAT
GGAAGTAGCA
CGTCTCACTA
GTCTCGTGCA
GATGGACAGC
ACCGCTGAGC
AATGGAAGCG
GGTAGGCCTT
TGGGGCAGCG
GCCAATAGCA
GCTTTGCTCC
TTCGCTTTCT
GGGCTCAGAG
GCTGGGAAGG
GGTGGGTCCG
GGGGCGGGCT
CAGGGGCGGG
CTCAGGGGCG
GGGCGGGCGC
CCGAAGGTCC
TCCGGAGGCC
CGGCATTCTG
CACGCTTCAA
AAGCGCACGT
CTGCCGCGCT
GTTCTCCTCT
TCCTCATCTC
CGGGCCTTTC
GACCTGCAGC
CCAAGCTTAC
CGCCACCATG
GTGAGCAAGG
GCGAGGAGCT
GTTCACCGGG
GTGGTGCCCA
TCCTGGTCGA
GCTGGACGGC
GACGTAAACG
GCCACAAGTT
CAGCGTGTCC
GGCGAGGGCG
AGGGCGATGC
CACCTACGGC
AAGCTGACCC
TGAAGTTCAT
CTGCACCACC
GGCAAGCTGC
CCGTGCCCTG
GCCCACCCTC
GTGACCACCC
TGACCTACGG
CGTGCAGTGC
TTCAGCCGCT
ACCCCGACCA
CATGAAGCAG
CACGACTTCT
TCAAGTCCGC
CATGCCCGAA
GGCTACGTCC
AGGAGCGCAC
CATCTTCTTC
AAGGACGACG
GCAACTACAA
GACCCGCGCC
GAGGTGAAGT
TCGAGGGCGA
CACCCTGGTG
AACCGCATCG
AGCTGAAGGG
CATCGACTTC
AAGGAGGACG
GCAACATCCT
GGGGCACAAG
CTGGAGTACA
ACTACAACAG
CCACAACGTC
TATATCATGG
CCGACAAGCA
GAAGAACGGC
ATCAAGGTGA
ACTTCAAGAT
CCGCCACAAC
ATCGAGGACG
GCAGCGTGCA
GCTCGCCGAC
CACTACCAGC
AGAACACCCC
CATCGGCGAC
GGCCCCGTGC
TGCTGCCCGA
CAACCACTAC
CTGAGCACCC
AGTCCGCCCT
GAGCAAAGAC
CCCAACGAGA
AGCGCGATCA
CATGGTCCTG
CTGGAGTTCG
TGACCGCCGC
CGGGATCACT
CTCGGCATGG
ACGAGCTGTA
CAAGGAATTT
GGCAGCGGCC
AGTGCACCAA
CTATGCCCTG
CTGAAACTGG
CTGGCGATGT
GGAAAGCAAC
CCCGGACCCA
TGACCGAGTA
CAAGCCCACG
GTGCGCCTCG
CCACCCGCGA
CGACGTCCCC
AGGGCCGTAC
GCACCCTCGC
CGCCGCGTTC
GCCGACTACC
CCGCCACGCG
CCACACCGTC
GATCCGGACC
GCCACATCGA
GCGGGTCACC
GAGCTGCAAG
AACTCTTCCT
CACGCGCGTC
GGGCTCGACA
TCGGCAAGGT
GTGGGTCGCG
GACGACGGCG
CCGCGGTGGC
GGTCTGGACC
ACGCCGGAGA
GCGTCGAAGC
GGGGGCGGTG
TTCGCCGAGA
TCGGCCCGCG
CATGGCCGAG
TTGAGCGGTT
CCCGGCTGGC
CGCGCAGCAA
CAGATGGAAG
GCCTCCTGGC
GCCGCACCGG
CCCAAGGAGC
CCGCGTGGTT
CCTGGCCACC
GTCGGCGTCT
CGCCCGACCA
CCAGGGCAAG
GGTCTGGGCA
GCGCCGTCGT
GCTCCCCGGA
GTGGAGGCGG
CCGAGCGCGC
CGGGGTGCCC
GCCTTCCTGG
AGACCTCCGC
GCCCCGCAAC
CTCCCCTTCT
ACGAGCGGCT
CGGCTTCACC
GTCACCGCCG
ACGTCGAGGT
GCCCGAAGGA
CCGCGCACCT
GGTGCATGAC
CCGCAAGCCC
GGTGCCTGAC
GCCCGCCCCA
CGACCCGCAG
CGCCCGACCG
AAAGGAGCGC
ACGACCCCAT
GCATCGGCGT
CTCGAGTCTA
GAGGGCCCGA
CGCGTCTGGA
ACAATCAACC
TCTGGATTAC
AAAATTTGTG
AAAGATTGAC
TGGTATTCTT
AACTATGTTG
CTCCTTTTAC
GCTATGTGGA
TACGCTGCTT
TAATGGCTTT
GTATCATGCT
ATTGCTTCCC
GTATGGCTTT
CATTTTCTCC
TCCTTGTATA
AATCCTGGTT
GCTGTCTCTT
TATGAGGAGT
TGTGGCCCGT
TGTCAGGCAA
CGTGGCGTGG
TGTGCACTGT
GTTTGCTGAC
GCAACCCCCA
CTGGTTGGGG
CATTGCCACC
ACCTGTCAGC
TCCTTTCCGG
GACTTTCGCT
TTCCCCCTCC
CTATTGCCAC
GGCGGAACTC
ATCGCCGCCT
GCCTTGCCCG
CTGCTGGACA
GGGGCTCGGC
TGTTGGGCAC
TGACAATTCC
GTGGTGTTGT
CGGGGAAGCT
GACGTCCTTT
CCATGGCTGC
TCGCCTGTGT
TGCCACCTGG
ATTCTGCGCG
GGACGTCCTT
CTGCTACGTC
CCTTCGGCCC
TCAATCCAGC
GGACCTTCCT
TCCCGCGGCC
TGCTGCCGGC
TCTGCGGCTT
CCGCGTCTTC
GCCTTCGCCC
TCAGACGAGT
CGGATCTCCC
TTTGGGCCGC
CTCCCCGCCT
GGAATTAATT
CTGCAGTCGA
GACCTAGAAA
AACATGGAGC
AATCACAAGT
AGCAATACAG
CAGCTATCAA
TGATTGTGCC
TGGCTAGAAG
CACAAGAGGA
GGAGGAGGTG
GGTTTTCCAG
TCACACCTCA
GGTACCTTTA
AGACCAATGA
CTTACAAGGC
AGCTGTAGAT
CTTAGCCACT
TTTTAAAAGA
AAAGAGGGGA
CTGGAAGGGC
TAATTCACTC
CCAACGAAGA
CAAGATATCC
TTGATCTGTG
GATCTACCAC
ACACAAGGCT
ACTTCCCTGA
TTAGCAGAAC
TACACACCAG
GGCCAGGGGT
CAGATATCCA
CTGACCTTTG
GATGGTGCTA
CAAGCTAGTA
CCAGTTGAGC
CAGATAAGGT
AGAAGAGGCC
AATAAAGGAG
AGAACACCAG
CTTGTTACAC
CCTGTGAGCC
TGCATGGGAT
GGATGACCCG
GAGAGAGAAG
TGTTAGAGTG
GAGGTTTGAC
AGCCGCCTAG
CATTTCATCA
CGTGGCCCGA
GAGCTGCATC
CGGAGTACTT
CAAGAACTGC
TGATATCGAG
CTTGCTACAA
GGGACTTTCC
GCTGGGGACT
TTCCAGGGAG
GCGTGGCCTG
GGCGGGACTG
GGGAGTGGCG
AGCCCTCAGA
TCCTGCATAT
AAGCAGCTGC
TTTTTGCCTG
TACTGGGTCT
CTCTGGTTAG
ACCAGATCTG
AGCCTGGGAG
CTCTCTGGCT
AACTAGGGAA
CCCACTGCTT
AAGCCTCAAT
AAAGCTTGCC
TTGAGTGCTT
CAAGTAGTGT
GTGCCCGTCT
GTTGTGTGAC
TCTGGTAACT
AGAGATCCCT
CAGACCCTTT
TAGTCAGTGT
GGAAAATCTC
TAGCAGTAGT
AGTTCATGTC
ATCTTATTAT
TCAGTATTTA
TAACTTGCAA
AGAAATGAAT
ATCAGAGAGT
GAGAGGCCTT
GACATTGCTA
GCGTTTACCG
TCGACCTCTA
GCTAGAGCTT
GGCGTAATCA
TGGTCATAGC
TGTTTCCTGT
GTGAAATTGT
TATCCGCTCA
CAATTCCACA
CAACATACGA
GCCGGAAGCA
TAAAGTGTAA
AGCCTGGGGT
GCCTAATGAG
TGAGCTAACT
CACATTAATT
GCGTTGCGCT
CACTGCCCGC
TTTCCAGTCG
GGAAACCTGT
CGTGCCAGCT
GCATTAATGA
ATCGGCCAAC
GCGCGGGGAG
AGGCGGTTTG
CGTATTGGGC
GCTCTTCCGC
TTCCTCGCTC
ACTGACTCGC
TGCGCTCGGT
CGTTCGGCTG
CGGCGAGCGG
TATCAGCTCA
CTCAAAGGCG
GTAATACGGT
TATCCACAGA
ATCAGGGGAT
AACGCAGGAA
AGAACATGTG
AGCAAAAGGC
CAGCAAAAGG
CCAGGAACCG
TAAAAAGGCC
GCGTTGCTGG
CGTTTTTCCA
TAGGCTCCGC
CCCCCTGACG
AGCATCACAA
AAATCGACGC
TCAAGTCAGA
GGTGGCGAAA
CCCGACAGGA
CTATAAAGAT
ACCAGGCGTT
TCCCCCTGGA
AGCTCCCTCG
TGCGCTCTCC
TGTTCCGACC
CTGCCGCTTA
CCGGATACCT
GTCCGCCTTT
CTCCCTTCGG
GAAGCGTGGC
GCTTTCTCAT
AGCTCACGCT
GTAGGTATCT
CAGTTCGGTG
TAGGTCGTTC
GCTCCAAGCT
GGGCTGTGTG
CACGAACCCC
CCGTTCAGCC
CGACCGCTGC
GCCTTATCCG
GTAACTATCG
TCTTGAGTCC
AACCCGGTAA
GACACGACTT
ATCGCCACTG
GCAGCAGCCA
CTGGTAACAG
GATTAGCAGA
GCGAGGTATG
TAGGCGGTGC
TACAGAGTTC
TTGAAGTGGT
GGCCTAACTA
CGGCTACACT
AGAAGAACAG
TATTTGGTAT
CTGCGCTCTG
CTGAAGCCAG
TTACCTTCGG
AAAAAGAGTT
GGTAGCTCTT
GATCCGGCAA
ACAAACCACC
GCTGGTAGCG
GTGGTTTTTT
TGTTTGCAAG
CAGCAGATTA
CGCGCAGAAA
AAAAGGATCT
CAAGAAGATC
CTTTGATCTT
TTCTACGGGG
TCTGACGCTC
AGTGGAACGA
AAACTCACGT
TAAGGGATTT
TGGTCATGAG
ATTATCAAAA
AGGATCTTCA
CCTAGATCCT
TTTAAATTAA
AAATGAAGTT
TTAAATCAAT
CTAAAGTATA
TATGAGTAAA
CTTGGTCTGA
CAGTTACCAA
TGCTTAATCA
GTGAGGCACC
TATCTCAGCG
ATCTGTCTAT
TTCGTTCATC
CATAGTTGCC
TGACTCCCCG
TCGTGTAGAT
AACTACGATA
CGGGAGGGCT
TACCATCTGG
CCCCAGTGCT
GCAATGATAC
CGCGAGACCC
ACGCTCACCG
GCTCCAGATT
TATCAGCAAT
AAACCAGCCA
GCCGGAAGGG
CCGAGCGCAG
AAGTGGTCCT
GCAACTTTAT
CCGCCTCCAT
CCAGTCTATT
AATTGTTGCC
GGGAAGCTAG
AGTAAGTAGT
TCGCCAGTTA
ATAGTTTGCG
CAACGTTGTT
GCCATTGCTA
CAGGCATCGT
GGTGTCACGC
TCGTCGTTTG
GTATGGCTTC
ATTCAGCTCC
GGTTCCCAAC
GATCAAGGCG
AGTTACATGA
TCCCCCATGT
TGTGCAAAAA
AGCGGTTAGC
TCCTTCGGTC
CTCCGATCGT
TGTCAGAAGT
AAGTTGGCCG
CAGTGTTATC
ACTCATGGTT
ATGGCAGCAC
TGCATAATTC
TCTTACTGTC
ATGCCATCCG
TAAGATGCTT
TTCTGTGACT
GGTGAGTACT
CAACCAAGTC
ATTCTGAGAA
TAGTGTATGC
GGCGACCGAG
TTGCTCTTGC
CCGGCGTCAA
TACGGGATAA
TACCGCGCCA
CATAGCAGAA
CTTTAAAAGT
GCTCATCATT
GGAAAACGTT
CTTCGGGGCG
AAAACTCTCA
AGGATCTTAC
CGCTGTTGAG
ATCCAGTTCG
ATGTAACCCA
CTCGTGCACC
CAACTGATCT
TCAGCATCTT
TTACTTTCAC
CAGCGTTTCT
GGGTGAGCAA
AAACAGGAAG
GCAAAATGCC
GCAAAAAAGG
GAATAAGGGC
GACACGGAAA
TGTTGAATAC
TCATACTCTT
CCTTTTTCAA
TATTATTGAA
GCATTTATCA
GGGTTATTGT
CTCATGAGCG
GATACATATT
TGAATGTATT
TAGAAAAATA
AACAAATAGG
GGTTCCGCGC
ACATTTCCCC
GAAAAGTGCC
ACCTGACGTC
GACGGATCGG
GAGATCAACT
TGTTTATTGC
AGCTTATAAT
GGTTACAAAT
AAAGCAATAG
CATCACAAAT
TTCACAAATA
AAGCATTTTT
TTCACTGCAT
TCTAGTTGTG
GTTTGTCCAA
ACTCATCAAT
GTATCTTATC
ATGTCTGGAT
CAACTGGATA
ACTCAAGCTA
ACCAAAATCA
TCCCAAACTT
CCCACCCCAT
ACCCTATTAC
CACTGCCAAT
TACCTGTGGT
TTCATTTACT
CTAAACCTGT
GATTCCTCTG
AATTATTTTC
ATTTTAAAGA
AATTGTATTT
GTTAAATATG
TACTACAAAC
TTAGTAGTTT
TTAAAGAAAT
TGTATTTGTT
AAATATGTAC
TACAAACTTA
GTAGT