名称: | LentiGuide_DKO |
---|---|
别称: | Plasmid #183193 |
基因长: | |
质粒宿主: | 哺乳细胞,慢病毒 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR |
片段物种: | 空载体 |
原核抗性: | Amp |
筛选标记: | Puro |
荧光标记: | |
启动子: | hU6, mU6 |
复制子: | pUC |
拷贝数: | |
感受态: | Stbl3 |
温度: | 30度 |
背景载体: | LentiGuide-Puro |
正向引物: | |
反向引物: | PLKO.1-R |
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。
TAACAAATTG
GCTGTGGTAT
ATAAAATTAT
TCATAATGAT
AGTAGGAGGC
TTGGTAGGTT
TAAGAATAGT
TTTTGCTGTA
CTTTCTATAG
TGAATAGAGT
TAGGCAGGGA
TATTCACCAT
TATCGTTTCA
GACCCACCTC
CCAACCCCGA
GGGGACCCAG
AGAGGGCCTA
TTTCCCATGA
TTCCTTCATA
TTTGCATATA
CGATACAAGG
CTGTTAGAGA
GATAATTAGA
ATTAATTTGA
CTGTAAACAC
AAAGATATTA
GTACAAAATA
CGTGACGTAG
AAAGTAATAA
TTTCTTGGGT
AGTTTGCAGT
TTTAAAATTA
TGTTTTAAAA
TGGACTATCA
TATGCTTACC
GTAACTTGAA
AGTATTTCGA
TTTCTTGGCT
TTATATATCT
TGTGGAAAGG
ACTCTAGAAC
CAGGATGGGC
ACCACCCGTT
TAAGAGCTAA
GCTGGAAACA
GCATAGCAAG
TTTAAATAAG
GCTAGTCCGT
TATCAACTTG
AAAAAGTGGC
ACCGAGTCGG
TGCTTTTTTT
GTACTGAGTC
GCCCAGTCTC
AGATAGATCC
GACGCCGCCA
TCTCTAGGCC
CGCGCCGGCC
CCCTCGCACA
GACTTGTGGG
AGAAGCTCGG
CTACTCCCCT
GCCCCGGTTA
ATTTGCATAT
AATATTTCCT
AGTAACTATA
GAGGCTTAAT
GTGCGATAAA
AGACAGATAA
TCTGTTCTTT
TTAATACTAG
CTACATTTTA
CATGATAGGC
TTGGATTTCT
ATAAGAGATA
CAAATACTAA
ATTATTATTT
TAAAAAACAG
CACAAAAGGA
AACTCACCCT
AACTGTAAAG
TAATTGTGTG
TTTTGAGACT
ATAAATATCC
CTTGGAGAAA
AGCCTTGTTC
ACCGGAGACG
GCTTCTTGCT
CTTTTTTGTA
CGTCTCTGTT
TTAGAGCTAG
AAATAGCAAG
TTAAAATAAG
GCTAGTCCGT
TATCAACTTG
AAAAAGTGGC
ACCGAGTCGG
TGCTTTTTTA
AGCTTGGCGT
AACTAGATCT
TGAGACAAAT
GGCAGTATTC
ATCCACAATT
TTAAAAGAAA
AGGGGGGATT
GGGGGGTACA
GTGCAGGGGA
AAGAATAGTA
GACATAATAG
CAACAGACAT
ACAAACTAAA
GAATTACAAA
AACAAATTAC
AAAAATTCAA
AATTTTCGGG
TTTATTACAG
GGACAGCAGA
GATCCACTTT
GGCGCCGGCT
CGAGGGGGCC
CGGGTGCAAA
GATGGATAAA
GTTTTAAACA
GAGAGGAATC
TTTGCAGCTA
ATGGACCTTC
TAGGTCTTGA
AAGGAGTGGG
AATTGGCTCC
GGTGCCCGTC
AGTGGGCAGA
GCGCACATCG
CCCACAGTCC
CCGAGAAGTT
GGGGGGAGGG
GTCGGCAATT
GATCCGGTGC
CTAGAGAAGG
TGGCGCGGGG
TAAACTGGGA
AAGTGATGTC
GTGTACTGGC
TCCGCCTTTT
TCCCGAGGGT
GGGGGAGAAC
CGTATATAAG
TGCAGTAGTC
GCCGTGAACG
TTCTTTTTCG
CAACGGGTTT
GCCGCCAGAA
CACAGGTAAG
TGCCGTGTGT
GGTTCCCGCG
GGCCTGGCCT
CTTTACGGGT
TATGGCCCTT
GCGTGCCTTG
AATTACTTCC
ACCTGGCTGC
AGTACGTGAT
TCTTGATCCC
GAGCTTCGGG
TTGGAAGTGG
GTGGGAGAGT
TCGAGGCCTT
GCGCTTAAGG
AGCCCCTTCG
CCTCGTGCTT
GAGTTGAGGC
CTGGCCTGGG
CGCTGGGGCC
GCCGCGTGCG
AATCTGGTGG
CACCTTCGCG
CCTGTCTCGC
TGCTTTCGAT
AAGTCTCTAG
CCATTTAAAA
TTTTTGATGA
CCTGCTGCGA
CGCTTTTTTT
CTGGCAAGAT
AGTCTTGTAA
ATGCGGGCCA
AGATCTGCAC
ACTGGTATTT
CGGTTTTTGG
GGCCGCGGGC
GGCGACGGGG
CCCGTGCGTC
CCAGCGCACA
TGTTCGGCGA
GGCGGGGCCT
GCGAGCGCGG
CCACCGAGAA
TCGGACGGGG
GTAGTCTCAA
GCTGGCCGGC
CTGCTCTGGT
GCCTGGCCTC
GCGCCGCCGT
GTATCGCCCC
GCCCTGGGCG
GCAAGGCTGG
CCCGGTCGGC
ACCAGTTGCG
TGAGCGGAAA
GATGGCCGCT
TCCCGGCCCT
GCTGCAGGGA
GCTCAAAATG
GAGGACGCGG
CGCTCGGGAG
AGCGGGCGGG
TGAGTCACCC
ACACAAAGGA
AAAGGGCCTT
TCCGTCCTCA
GCCGTCGCTT
CATGTGACTC
CACGGAGTAC
CGGGCGCCGT
CCAGGCACCT
CGATTAGTTC
TCGAGCTTTT
GGAGTACGTC
GTCTTTAGGT
TGGGGGGAGG
GGTTTTATGC
GATGGAGTTT
CCCCACACTG
AGTGGGTGGA
GACTGAAGTT
AGGCCAGCTT
GGCACTTGAT
GTAATTCTCC
TTGGAATTTG
CCCTTTTTGA
GTTTGGATCT
TGGTTCATTC
TCAAGCCTCA
GACAGTGGTT
CAAAGTTTTT
TTCTTCCATT
TCAGGTGTCG
TGACGTACGG
CCACCATGAC
CGAGTACAAG
CCCACGGTGC
GCCTCGCCAC
CCGCGACGAC
GTCCCCAGGG
CCGTACGCAC
CCTCGCCGCC
GCGTTCGCCG
ACTACCCCGC
CACGCGCCAC
ACCGTCGATC
CGGACCGCCA
CATCGAGCGG
GTCACCGAGC
TGCAAGAACT
CTTCCTCACG
CGCGTCGGGC
TCGACATCGG
CAAGGTGTGG
GTCGCGGACG
ACGGCGCCGC
CGTGGCGGTC
TGGACCACGC
CGGAGAGCGT
CGAAGCGGGG
GCGGTGTTCG
CCGAGATCGG
CCCGCGCATG
GCCGAGTTGA
GCGGTTCCCG
GCTGGCCGCG
CAGCAACAGA
TGGAAGGCCT
CCTGGCGCCG
CACCGGCCCA
AGGAGCCCGC
GTGGTTCCTG
GCCACCGTCG
GAGTCTCGCC
CGACCACCAG
GGCAAGGGTC
TGGGCAGCGC
CGTCGTGCTC
CCCGGAGTGG
AGGCGGCCGA
GCGCGCCGGG
GTGCCCGCCT
TCCTGGAGAC
CTCCGCGCCC
CGCAACCTCC
CCTTCTACGA
GCGGCTCGGC
TTCACCGTCA
CCGCCGACGT
CGAGGTGCCC
GAAGGACCGC
GCACCTGGTG
CATGACCCGC
AAGCCCGGTG
CCTGAACGCG
TTAAGTCGAC
AATCAACCTC
TGGATTACAA
AATTTGTGAA
AGATTGACTG
GTATTCTTAA
CTATGTTGCT
CCTTTTACGC
TATGTGGATA
CGCTGCTTTA
ATGCCTTTGT
ATCATGCTAT
TGCTTCCCGT
ATGGCTTTCA
TTTTCTCCTC
CTTGTATAAA
TCCTGGTTGC
TGTCTCTTTA
TGAGGAGTTG
TGGCCCGTTG
TCAGGCAACG
TGGCGTGGTG
TGCACTGTGT
TTGCTGACGC
AACCCCCACT
GGTTGGGGCA
TTGCCACCAC
CTGTCAGCTC
CTTTCCGGGA
CTTTCGCTTT
CCCCCTCCCT
ATTGCCACGG
CGGAACTCAT
CGCCGCCTGC
CTTGCCCGCT
GCTGGACAGG
GGCTCGGCTG
TTGGGCACTG
ACAATTCCGT
GGTGTTGTCG
GGGAAATCAT
CGTCCTTTCC
TTGGCTGCTC
GCCTGTGTTG
CCACCTGGAT
TCTGCGCGGG
ACGTCCTTCT
GCTACGTCCC
TTCGGCCCTC
AATCCAGCGG
ACCTTCCTTC
CCGCGGCCTG
CTGCCGGCTC
TGCGGCCTCT
TCCGCGTCTT
CGCCTTCGCC
CTCAGACGAG
TCGGATCTCC
CTTTGGGCCG
CCTCCCCGCG
TCGACTTTAA
GACCAATGAC
TTACAAGGCA
GCTGTAGATC
TTAGCCACTT
TTTAAAAGAA
AAGGGGGGAC
TGGAAGGGCT
AATTCACTCC
CAACGAAGAC
AAGATCTGCT
TTTTGCTTGT
ACTGGGTCTC
TCTGGTTAGA
CCAGATCTGA
GCCTGGGAGC
TCTCTGGCTA
ACTAGGGAAC
CCACTGCTTA
AGCCTCAATA
AAGCTTGCCT
TGAGTGCTTC
AAGTAGTGTG
TGCCCGTCTG
TTGTGTGACT
CTGGTAACTA
GAGATCCCTC
AGACCCTTTT
AGTCAGTGTG
GAAAATCTCT
AGCAGTACGT
ATAGTAGTTC
ATGTCATCTT
ATTATTCAGT
ATTTATAACT
TGCAAAGAAA
TGAATATCAG
AGAGTGAGAG
GAACTTGTTT
ATTGCAGCTT
ATAATGGTTA
CAAATAAAGC
AATAGCATCA
CAAATTTCAC
AAATAAAGCA
TTTTTTTCAC
TGCATTCTAG
TTGTGGTTTG
TCCAAACTCA
TCAATGTATC
TTATCATGTC
TGGCTCTAGC
TATCCCGCCC
CTAACTCCGC
CCATCCCGCC
CCTAACTCCG
CCCAGTTCCG
CCCATTCTCC
GCCCCATGGC
TGACTAATTT
TTTTTATTTA
TGCAGAGGCC
GAGGCCGCCT
CGGCCTCTGA
GCTATTCCAG
AAGTAGTGAG
GAGGCTTTTT
TGGAGGCCTA
GGGACGTACC
CAATTCGCCC
TATAGTGAGT
CGTATTACGC
GCGCTCACTG
GCCGTCGTTT
TACAACGTCG
TGACTGGGAA
AACCCTGGCG
TTACCCAACT
TAATCGCCTT
GCAGCACATC
CCCCTTTCGC
CAGCTGGCGT
AATAGCGAAG
AGGCCCGCAC
CGATCGCCCT
TCCCAACAGT
TGCGCAGCCT
GAATGGCGAA
TGGGACGCGC
CCTGTAGCGG
CGCATTAAGC
GCGGCGGGTG
TGGTGGTTAC
GCGCAGCGTG
ACCGCTACAC
TTGCCAGCGC
CCTAGCGCCC
GCTCCTTTCG
CTTTCTTCCC
TTCCTTTCTC
GCCACGTTCG
CCGGCTTTCC
CCGTCAAGCT
CTAAATCGGG
GGCTCCCTTT
AGGGTTCCGA
TTTAGTGCTT
TACGGCACCT
CGACCCCAAA
AAACTTGATT
AGGGTGATGG
TTCACGTAGT
GGGCCATCGC
CCTGATAGAC
GGTTTTTCGC
CCTTTGACGT
TGGAGTCCAC
GTTCTTTAAT
AGTGGACTCT
TGTTCCAAAC
TGGAACAACA
CTCAACCCTA
TCTCGGTCTA
TTCTTTTGAT
TTATAAGGGA
TTTTGCCGAT
TTCGGCCTAT
TGGTTAAAAA
ATGAGCTGAT
TTAACAAAAA
TTTAACGCGA
ATTTTAACAA
AATATTAACG
CTTACAATTT
AGGTGGCACT
TTTCGGGGAA
ATGTGCGCGG
AACCCCTATT
TGTTTATTTT
TCTAAATACA
TTCAAATATG
TATCCGCTCA
TGAGACAATA
ACCCTGATAA
ATGCTTCAAT
AATATTGAAA
AAGGAAGAGT
ATGAGTATTC
AACATTTCCG
TGTCGCCCTT
ATTCCCTTTT
TTGCGGCATT
TTGCCTTCCT
GTTTTTGCTC
ACCCAGAAAC
GCTGGTGAAA
GTAAAAGATG
CTGAAGATCA
GTTGGGTGCA
CGAGTGGGTT
ACATCGAACT
GGATCTCAAC
AGCGGTAAGA
TCCTTGAGAG
TTTTCGCCCC
GAAGAACGTT
TTCCAATGAT
GAGCACTTTT
AAAGTTCTGC
TATGTGGCGC
GGTATTATCC
CGTATTGACG
CCGGGCAAGA
GCAACTCGGT
CGCCGCATAC
ACTATTCTCA
GAATGACTTG
GTTGAGTACT
CACCAGTCAC
AGAAAAGCAT
CTTACGGATG
GCATGACAGT
AAGAGAATTA
TGCAGTGCTG
CCATAACCAT
GAGTGATAAC
ACTGCGGCCA
ACTTACTTCT
GACAACGATC
GGAGGACCGA
AGGAGCTAAC
CGCTTTTTTG
CACAACATGG
GGGATCATGT
AACTCGCCTT
GATCGTTGGG
AACCGGAGCT
GAATGAAGCC
ATACCAAACG
ACGAGCGTGA
CACCACGATG
CCTGTAGCAA
TGGCAACAAC
GTTGCGCAAA
CTATTAACTG
GCGAACTACT
TACTCTAGCT
TCCCGGCAAC
AATTAATAGA
CTGGATGGAG
GCGGATAAAG
TTGCAGGACC
ACTTCTGCGC
TCGGCCCTTC
CGGCTGGCTG
GTTTATTGCT
GATAAATCTG
GAGCCGGTGA
GCGTGGGTCT
CGCGGTATCA
TTGCAGCACT
GGGGCCAGAT
GGTAAGCCCT
CCCGTATCGT
AGTTATCTAC
ACGACGGGGA
GTCAGGCAAC
TATGGATGAA
CGAAATAGAC
AGATCGCTGA
GATAGGTGCC
TCACTGATTA
AGCATTGGTA
ACTGTCAGAC
CAAGTTTACT
CATATATACT
TTAGATTGAT
TTAAAACTTC
ATTTTTAATT
TAAAAGGATC
TAGGTGAAGA
TCCTTTTTGA
TAATCTCATG
ACCAAAATCC
CTTAACGTGA
GTTTTCGTTC
CACTGAGCGT
CAGACCCCGT
AGAAAAGATC
AAAGGATCTT
CTTGAGATCC
TTTTTTTCTG
CGCGTAATCT
GCTGCTTGCA
AACAAAAAAA
CCACCGCTAC
CAGCGGTGGT
TTGTTTGCCG
GATCAAGAGC
TACCAACTCT
TTTTCCGAAG
GTAACTGGCT
TCAGCAGAGC
GCAGATACCA
AATACTGTTC
TTCTAGTGTA
GCCGTAGTTA
GGCCACCACT
TCAAGAACTC
TGTAGCACCG
CCTACATACC
TCGCTCTGCT
AATCCTGTTA
CCAGTGGCTG
CTGCCAGTGG
CGATAAGTCG
TGTCTTACCG
GGTTGGACTC
AAGACGATAG
TTACCGGATA
AGGCGCAGCG
GTCGGGCTGA
ACGGGGGGTT
CGTGCACACA
GCCCAGCTTG
GAGCGAACGA
CCTACACCGA
ACTGAGATAC
CTACAGCGTG
AGCTATGAGA
AAGCGCCACG
CTTCCCGAAG
GGAGAAAGGC
GGACAGGTAT
CCGGTAAGCG
GCAGGGTCGG
AACAGGAGAG
CGCACGAGGG
AGCTTCCAGG
GGGAAACGCC
TGGTATCTTT
ATAGTCCTGT
CGGGTTTCGC
CACCTCTGAC
TTGAGCGTCG
ATTTTTGTGA
TGCTCGTCAG
GGGGGCGGAG
CCTATGGAAA
AACGCCAGCA
ACGCGGCCTT
TTTACGGTTC
CTGGCCTTTT
GCTGGCCTTT
TGCTCACATG
TTCTTTCCTG
CGTTATCCCC
TGATTCTGTG
GATAACCGTA
TTACCGCCTT
TGAGTGAGCT
GATACCGCTC
GCCGCAGCCG
AACGACCGAG
CGCAGCGAGT
CAGTGAGCGA
GGAAGCGGAA
GAGCGCCCAA
TACGCAAACC
GCCTCTCCCC
GCGCGTTGGC
CGATTCATTA
ATGCAGCTGG
CACGACAGGT
TTCCCGACTG
GAAAGCGGGC
AGTGAGCGCA
ACGCAATTAA
TGTGAGTTAG
CTCACTCATT
AGGCACCCCA
GGCTTTACAC
TTTATGCTTC
CGGCTCGTAT
GTTGTGTGGA
ATTGTGAGCG
GATAACAATT
TCACACAGGA
AACAGCTATG
ACCATGATTA
CGCCAAGCGC
GCAATTAACC
CTCACTAAAG
GGAACAAAAG
CTGGAGCTGC
AAGCTTAATG
TAGTCTTATG
CAATACTCTT
GTAGTCTTGC
AACATGGTAA
CGATGAGTTA
GCAACATGCC
TTACAAGGAG
AGAAAAAGCA
CCGTGCATGC
CGATTGGTGG
AAGTAAGGTG
GTACGATCGT
GCCTTATTAG
GAAGGCAACA
GACGGGTCTG
ACATGGATTG
GACGAACCAC
TGAATTGCCG
CATTGCAGAG
ATATTGTATT
TAAGTGCCTA
GCTCGATACA
TAAACGGGTC
TCTCTGGTTA
GACCAGATCT
GAGCCTGGGA
GCTCTCTGGC
TAACTAGGGA
ACCCACTGCT
TAAGCCTCAA
TAAAGCTTGC
CTTGAGTGCT
TCAAGTAGTG
TGTGCCCGTC
TGTTGTGTGA
CTCTGGTAAC
TAGAGATCCC
TCAGACCCTT
TTAGTCAGTG
TGGAAAATCT
CTAGCAGTGG
CGCCCGAACA
GGGACTTGAA
AGCGAAAGGG
AAACCAGAGG
AGCTCTCTCG
ACGCAGGACT
CGGCTTGCTG
AAGCGCGCAC
GGCAAGAGGC
GAGGGGCGGC
GACTGGTGAG
TACGCCAAAA
ATTTTGACTA
GCGGAGGCTA
GAAGGAGAGA
GATGGGTGCG
AGAGCGTCAG
TATTAAGCGG
GGGAGAATTA
GATCGCGATG
GGAAAAAATT
CGGTTAAGGC
CAGGGGGAAA
GAAAAAATAT
AAATTAAAAC
ATATAGTATG
GGCAAGCAGG
GAGCTAGAAC
GATTCGCAGT
TAATCCTGGC
CTGTTAGAAA
CATCAGAAGG
CTGTAGACAA
ATACTGGGAC
AGCTACAACC
ATCCCTTCAG
ACAGGATCAG
AAGAACTTAG
ATCATTATAT
AATACAGTAG
CAACCCTCTA
TTGTGTGCAT
CAAAGGATAG
AGATAAAAGA
CACCAAGGAA
GCTTTAGACA
AGATAGAGGA
AGAGCAAAAC
AAAAGTAAGA
CCACCGCACA
GCAAGCGGCC
GCTGATCTTC
AGACCTGGAG
GAGGAGATAT
GAGGGACAAT
TGGAGAAGTG
AATTATATAA
ATATAAAGTA
GTAAAAATTG
AACCATTAGG
AGTAGCACCC
ACCAAGGCAA
AGAGAAGAGT
GGTGCAGAGA
GAAAAAAGAG
CAGTGGGAAT
AGGAGCTTTG
TTCCTTGGGT
TCTTGGGAGC
AGCAGGAAGC
ACTATGGGCG
CAGCGTCAAT
GACGCTGACG
GTACAGGCCA
GACAATTATT
GTCTGGTATA
GTGCAGCAGC
AGAACAATTT
GCTGAGGGCT
ATTGAGGCGC
AACAGCATCT
GTTGCAACTC
ACAGTCTGGG
GCATCAAGCA
GCTCCAGGCA
AGAATCCTGG
CTGTGGAAAG
ATACCTAAAG
GATCAACAGC
TCCTGGGGAT
TTGGGGTTGC
TCTGGAAAAC
TCATTTGCAC
CACTGCTGTG
CCTTGGAATG
CTAGTTGGAG
TAATAAATCT
CTGGAACAGA
TTTGGAATCA
CACGACCTGG
ATGGAGTGGG
ACAGAGAAAT
TAACAATTAC
ACAAGCTTAA
TACACTCCTT
AATTGAAGAA
TCGCAAAACC
AGCAAGAAAA
GAATGAACAA
GAATTATTGG
AATTAGATAA
ATGGGCAAGT
TTGTGGAATT
GGTTTAACA