名称: | pX330S-2-PITCh |
---|---|
别称: | Plasmid#63670 |
基因长: | |
质粒宿主: | 哺乳细胞 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR |
片段物种: | 空载体 |
原核抗性: | Spe |
筛选标记: | |
荧光标记: | |
启动子: | U6 |
复制子: | pUC |
拷贝数: | |
感受态: | DH5a |
温度: | 37度 |
背景载体: | |
正向引物: | U6-F:GAGGGCCTATTTCCCATGA |
反向引物: | BGH:TAGAAGGCACAGTCGAGG |
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。
TGAGACCCGT
TACATAACTT
ACGGTAAATG
GCCCGCCTGG
CTGACCGCCC
AACGACCCCC
GCCCATTGAC
GTCAATAGTA
ACGCCAATAG
GGACTTTCCA
TTGACGTCAA
TGGGTGGAGT
ATTTACGGTA
AACTGCCCAC
TTGGCAGTAC
ATCAAGTGTA
TCATATGCCA
AGTACGCCCC
CTATTGACGT
CAATGACGGT
AAATGGCCCG
CCTGGCATTG
TGCCCAGTAC
ATGACCTTAT
GGGACTTTCC
TACTTGGCAG
TACATCTACG
TATTAGTCAT
CGCTATTACC
ATGGTCGAGG
TGAGCCCCAC
GTTCTGCTTC
ACTCTCCCCA
TCTCCCCCCC
CTCCCCACCC
CCAATTTTGT
ATTTATTTAT
TTTTTAATTA
TTTTGTGCAG
CGATGGGGGC
GGGGGGGGGG
GGGGGCGCGC
GCCrGGsGGG
GsGGGGsGGG
GGGGGGGGsG
GGGsGGGGsG
rGGCGGAGAG
GTGCGGCGGC
AGCCAATCAG
AGCGGCGCGC
TCCGAAAGTT
TCCTTTTATG
GCGAGGCGGC
GGCGGCGGCG
GCCCTATAAA
AAGCGAAGCG
CGCGGCGGGC
GGGAGTCGCT
GCGCGCTGCC
TTCGCCCCGT
GCCCCGCTCC
GCCGCCGCCT
CGCGCCGCCC
GCCCCGGCTC
TGACTGACCG
CGTTACTCCC
ACAGGTGAGC
GGGCGGGACG
GCCCTTCTCC
TCCGGGCTGT
AATTAGCTGA
GCAAGAGGTA
AGGGTTTAAG
GGATGGTTGG
TTGGTGGGGT
ATTAATGTTT
AATTACCTGG
AGCACCTGCC
TGAAATCACT
TTTTTTCAGG
TTGGACCGGT
GCCACCATGG
ACTATAAGGA
CCACGACGGA
GACTACAAGG
ATCATGATAT
TGATTACAAA
GACGATGACG
ATAAGATGGC
CCCAAAGAAG
AAGCGGAAGG
TCGGTATCCA
CGGAGTCCCA
GCAGCCGACA
AGAAGTACAG
CATCGGCCTG
GACATCGGCA
CCAACTCTGT
GGGCTGGGCC
GTGATCACCG
ACGAGTACAA
GGTGCCCAGC
AAGAAATTCA
AGGTGCTGGG
CAACACCGAC
CGGCACAGCA
TCAAGAAGAA
CCTGATCGGA
GCCCTGCTGT
TCGACAGCGG
CGAAACAGCC
GAGGCCACCC
GGCTGAAGAG
AACCGCCAGA
AGAAGATACA
CCAGACGGAA
GAACCGGATC
TGCTATCTGC
AAGAGATCTT
CAGCAACGAG
ATGGCCAAGG
TGGACGACAG
CTTCTTCCAC
AGACTGGAAG
AGTCCTTCCT
GGTGGAAGAG
GATAAGAAGC
ACGAGCGGCA
CCCCATCTTC
GGCAACATCG
TGGACGAGGT
GGCCTACCAC
GAGAAGTACC
CCACCATCTA
CCACCTGAGA
AAGAAACTGG
TGGACAGCAC
CGACAAGGCC
GACCTGCGGC
TGATCTATCT
GGCCCTGGCC
CACATGATCA
AGTTCCGGGG
CCACTTCCTG
ATCGAGGGCG
ACCTGAACCC
CGACAACAGC
GACGTTGACA
AGCTGTTCAT
CCAGCTGGTG
CAGACCTACA
ACCAGCTGTT
CGAGGAAAAC
CCCATCAACG
CCAGCGGCGT
GGACGCCAAG
GCCATCCTGT
CTGCCAGACT
GAGCAAGAGC
AGACGGCTGG
AAAATCTGAT
CGCCCAGCTG
CCCGGCGAGA
AGAAGAATGG
CCTGTTCGGA
AACCTGATTG
CCCTGAGCCT
GGGCCTGACC
CCCAACTTCA
AGAGCAACTT
CGACCTGGCC
GAGGATGCCA
AACTGCAGCT
GAGCAAGGAC
ACCTACGACG
ACGACCTGGA
CAACCTGCTG
GCCCAGATCG
GCGACCAGTA
CGCCGACCTG
TTTCTGGCCG
CCAAGAACCT
GTCCGACGCC
ATCCTGCTGA
GCGACATCCT
GAGAGTGAAC
ACCGAGATCA
CCAAGGCCCC
CCTGAGCGCC
TCTATGATCA
AGAGATACGA
CGAGCACCAC
CAGGACCTGA
CCCTGCTGAA
AGCTCTCGTG
CGGCAGCAGC
TGCCTGAGAA
GTACAAAGAG
ATTTTCTTCG
ACCAGAGCAA
GAACGGCTAC
GCCGGCTACA
TTGACGGCGG
AGCCAGCCAG
GAAGAGTTCT
ACAAGTTCAT
CAAGCCCATC
CTGGAAAAGA
TGGACGGCAC
CGAGGAACTG
CTCGTGAAGC
TGAACAGAGA
GGACCTGCTG
CGGAAGCAGC
GGACCTTCGA
CAACGGCAGC
ATCCCCCACC
AGATCCACCT
GGGAGAGCTG
CACGCCATTC
TGCGGCGGCA
GGAAGATTTT
TACCCATTCC
TGAAGGACAA
CCGGGAAAAG
ATCGAGAAGA
TCCTGACCTT
CCGCATCCCC
TACTACGTGG
GCCCTCTGGC
CAGGGGAAAC
AGCAGATTCG
CCTGGATGAC
CAGAAAGAGC
GAGGAAACCA
TCACCCCCTG
GAACTTCGAG
GAAGTGGTGG
ACAAGGGCGC
TTCCGCCCAG
AGCTTCATCG
AGCGGATGAC
CAACTTCGAT
AAGAACCTGC
CCAACGAGAA
GGTGCTGCCC
AAGCACAGCC
TGCTGTACGA
GTACTTCACC
GTGTATAACG
AGCTGACCAA
AGTGAAATAC
GTGACCGAGG
GAATGAGAAA
GCCCGCCTTC
CTGAGCGGCG
AGCAGAAAAA
GGCCATCGTG
GACCTGCTGT
TCAAGACCAA
CCGGAAAGTG
ACCGTGAAGC
AGCTGAAAGA
GGACTACTTC
AAGAAAATCG
AGTGCTTCGA
CTCCGTGGAA
ATCTCCGGCG
TGGAAGATCG
GTTCAACGCC
TCCCTGGGCA
CATACCACGA
TCTGCTGAAA
ATTATCAAGG
ACAAGGACTT
CCTGGACAAT
GAGGAAAACG
AGGACATTCT
GGAAGATATC
GTGCTGACCC
TGACACTGTT
TGAGGACAGA
GAGATGATCG
AGGAACGGCT
GAAAACCTAT
GCCCACCTGT
TCGACGACAA
AGTGATGAAG
CAGCTGAAGC
GGCGGAGATA
CACCGGCTGG
GGCAGGCTGA
GCCGGAAGCT
GATCAACGGC
ATCCGGGACA
AGCAGTCCGG
CAAGACAATC
CTGGATTTCC
TGAAGTCCGA
CGGCTTCGCC
AACAGAAACT
TCATGCAGCT
GATCCACGAC
GACAGCCTGA
CCTTTAAAGA
GGACATCCAG
AAAGCCCAGG
TGTCCGGCCA
GGGCGATAGC
CTGCACGAGC
ACATTGCCAA
TCTGGCCGGC
AGCCCCGCCA
TTAAGAAGGG
CATCCTGCAG
ACAGTGAAGG
TGGTGGACGA
GCTCGTGAAA
GTGATGGGCC
GGCACAAGCC
CGAGAACATC
GTGATCGAAA
TGGCCAGAGA
GAACCAGACC
ACCCAGAAGG
GACAGAAGAA
CAGCCGCGAG
AGAATGAAGC
GGATCGAAGA
GGGCATCAAA
GAGCTGGGCA
GCCAGATCCT
GAAAGAACAC
CCCGTGGAAA
ACACCCAGCT
GCAGAACGAG
AAGCTGTACC
TGTACTACCT
GCAGAATGGG
CGGGATATGT
ACGTGGACCA
GGAACTGGAC
ATCAACCGGC
TGTCCGACTA
CGATGTGGAC
CATATCGTGC
CTCAGAGCTT
TCTGAAGGAC
GACTCCATCG
ACAACAAGGT
GCTGACCAGA
AGCGACAAGA
ACCGGGGCAA
GAGCGACAAC
GTGCCCTCCG
AAGAGGTCGT
GAAGAAGATG
AAGAACTACT
GGCGGCAGCT
GCTGAACGCC
AAGCTGATTA
CCCAGAGAAA
GTTCGACAAT
CTGACCAAGG
CCGAGAGAGG
CGGCCTGAGC
GAACTGGATA
AGGCCGGCTT
CATCAAGAGA
CAGCTGGTGG
AAACCCGGCA
GATCACAAAG
CACGTGGCAC
AGATCCTGGA
CTCCCGGATG
AACACTAAGT
ACGACGAGAA
TGACAAGCTG
ATCCGGGAAG
TGAAAGTGAT
CACCCTGAAG
TCCAAGCTGG
TGTCCGATTT
CCGGAAGGAT
TTCCAGTTTT
ACAAAGTGCG
CGAGATCAAC
AACTACCACC
ACGCCCACGA
CGCCTACCTG
AACGCCGTCG
TGGGAACCGC
CCTGATCAAA
AAGTACCCTA
AGCTGGAAAG
CGAGTTCGTG
TACGGCGACT
ACAAGGTGTA
CGACGTGCGG
AAGATGATCG
CCAAGAGCGA
GCAGGAAATC
GGCAAGGCTA
CCGCCAAGTA
CTTCTTCTAC
AGCAACATCA
TGAACTTTTT
CAAGACCGAG
ATTACCCTGG
CCAACGGCGA
GATCCGGAAG
CGGCCTCTGA
TCGAGACAAA
CGGCGAAACC
GGGGAGATCG
TGTGGGATAA
GGGCCGGGAT
TTTGCCACCG
TGCGGAAAGT
GCTGAGCATG
CCCCAAGTGA
ATATCGTGAA
AAAGACCGAG
GTGCAGACAG
GCGGCTTCAG
CAAAGAGTCT
ATCCTGCCCA
AGAGGAACAG
CGATAAGCTG
ATCGCCAGAA
AGAAGGACTG
GGACCCTAAG
AAGTACGGCG
GCTTCGACAG
CCCCACCGTG
GCCTATTCTG
TGCTGGTGGT
GGCCAAAGTG
GAAAAGGGCA
AGTCCAAGAA
ACTGAAGAGT
GTGAAAGAGC
TGCTGGGGAT
CACCATCATG
GAAAGAAGCA
GCTTCGAGAA
GAATCCCATC
GACTTTCTGG
AAGCCAAGGG
CTACAAAGAA
GTGAAAAAGG
ACCTGATCAT
CAAGCTGCCT
AAGTACTCCC
TGTTCGAGCT
GGAAAACGGC
CGGAAGAGAA
TGCTGGCCTC
TGCCGGCGAA
CTGCAGAAGG
GAAACGAACT
GGCCCTGCCC
TCCAAATATG
TGAACTTCCT
GTACCTGGCC
AGCCACTATG
AGAAGCTGAA
GGGCTCCCCC
GAGGATAATG
AGCAGAAACA
GCTGTTTGTG
GAACAGCACA
AGCACTACCT
GGACGAGATC
ATCGAGCAGA
TCAGCGAGTT
CTCCAAGAGA
GTGATCCTGG
CCGACGCTAA
TCTGGACAAA
GTGCTGTCCG
CCTACAACAA
GCACCGGGAT
AAGCCCATCA
GAGAGCAGGC
CGAGAATATC
ATCCACCTGT
TTACCCTGAC
CAATCTGGGA
GCCCCTGCCG
CCTTCAAGTA
CTTTGACACC
ACCATCGACC
GGAAGAGGTA
CACCAGCACC
AAAGAGGTGC
TGGACGCCAC
CCTGATCCAC
CAGAGCATCA
CCGGCCTGTA
CGAGACACGG
ATCGACCTGT
CTCAGCTGGG
AGGCGACAAA
AGGCCGGCGG
CCACGAAAAA
GGCCGGCCAG
GCAAAAAAGA
AAAAGTAAGA
ATTCCTAGAG
CTCGCTGATC
AGCCTCGACT
GTGCCTTCTA
GTTGCCAGCC
ATCTGTTGTT
TGCCCCTCCC
CCGTGCCTTC
CTTGACCCTG
GAAGGTGCCA
CTCCCACTGT
CCTTTCCTAA
TAAAATGAGG
AAATTGCATC
GCATTGTCTG
AGTAGGTGTC
ATTCTATTCT
GGGGGGTGGG
GTGGGGCAGG
ACAGCAAGGG
GGAGGATTGG
GAAGAGAATA
GCAGGCATGC
TGGGGAGCGG
CCGCAGGAAC
CCCTAGTGAT
GGAGTTGGCC
ACTCCCTCTC
TGCGCGCTCG
CTCGCTCACT
GAGGCCGGGC
GACCAAAGGT
CGCCCGACGC
CCGGGCTTTG
CCCGGGCGGC
CTCAGTGAGC
GAGCGAGCGC
GCAGCTGCCT
GCAGGGGCGC
CTGATGCGGT
ATTTTCTCCT
TACGCATCTG
TGCGGTATTT
CACACCGCAT
ACGTCAAAGC
AACCATAGTA
CGCGCCCTGT
AGCGGCGCAT
TAAGCGCGGC
GGGTGTGGTG
GTTACGCGCA
GCGTGACCGC
TACACTTGCC
AGCGCCTTAG
CGCCCGCTCC
TTTCGCTTTC
TTCCCTTCCT
TTCTCGCCAC
GTTCGCCGGC
TTTCCCCGTC
AAGCTCTAAA
TCGGGGGCTC
CCTTTAGGGT
TCCGATTTAG
TGCTTTACGG
CACCTCGACC
CCAAAAAACT
TGATTTGGGT
GATGGTTCAC
GTAGTGGGCC
ATCGCCCTGA
TAGACGGTTT
TTCGCCCTTT
GACGTTGGAG
TCCACGTTCT
TTAATAGTGG
ACTCTTGTTC
CAAACTGGAA
CAACACTCAA
CTCTATCTCG
GGCTATTCTT
TTGATTTATA
AGGGATTTTG
CCGATTTCGG
TCTATTGGTT
AAAAAATGAG
CTGATTTAAC
AAAAATTTAA
CGCGAATTTT
AACAAAATAT
TAACGTTTAC
AATTTTATGG
TGCACTCTCA
GTACAATCTG
CTCTGATGCC
GCATAGTTAA
GCCAGCCCCG
ACACCCGCCA
ACACCCGCTG
ACGCGCCCTG
ACGGGCTTGT
CTGCTCCCGG
CATCCGCTTA
CAGACAAGCT
GTGACCGTCT
CCGGGAGCTG
CATGTGTCAG
AGGTTTTCAC
CGTCATCACC
GAAACGCGCG
AGACGAAAGG
GCCTCGTGAT
ACGCCTATTT
TTATAGGTTA
ATGTCATGAT
AATAATGGTT
TCTTAGACGT
CAGGTGGCAC
TTCCAGCCAG
GACAGAAATG
CCTCGACTTC
GCTGCTACCC
AAGGTTGCCG
GGTGACGCAC
ACCGTGGAAA
CGGATGAAGG
CACGAACCCA
GTGGACATAA
GCCTGTTCGG
TTCGTAAGCT
GTAATGCAAG
TAGCGTATGC
GCTCACGCAA
CTGGTCCAGA
ACCTTGACCG
AACGCAGCGG
TGGTAACGGC
GCAGTGGCGG
TTTTCATGGC
TTGTTATGAC
TGTTTTTTTG
GGGTACAGTC
TATGCCTCGG
GCATCCAAGC
AGCAAGCGCG
TTACGCCGTG
GGTCGATGTT
TGATGTTATG
GAGCAGCAAC
GATGTTACGC
AGCAGGGCAG
TCGCCCTAAA
ACAAAGTTAA
ACATTATGAG
GGAAGCGGTG
ATCGCCGAAG
TATCGACTCA
ACTATCAGAG
GTAGTTGGCG
TCATCGAGCG
CCATCTCGAA
CCGACGTTGC
TGGCCGTACA
TTTGTACGGC
TCCGCAGTGG
ATGGCGGCCT
GAAGCCACAC
AGTGATATTG
ATTTGCTGGT
TACGGTGACC
GTAAGGCTTG
ATGAAACAAC
GCGGCGAGCT
TTGATCAACG
ACCTTTTGGA
AACTTCGGCT
TCCCCTGGAG
AGAGCGAGAT
TCTCCGCGCT
GTAGAAGTCA
CCATTGTTGT
GCACGACGAC
ATCATTCCGT
GGCGTTATCC
AGCTAAGCGC
GAACTGCAAT
TTGGAGAATG
GCAGCGCAAT
GACATTCTTG
CAGGTATCTT
CGAGCCAGCC
ACGATCGACA
TTGATCTGGC
TATCTTGCTG
ACAAAAGCAA
GAGAACATAG
CGTTGCCTTG
GTAGGTCCAG
CGGCGGAGGA
ACTCTTTGAT
CCGGTTCCTG
AACAGGATCT
ATTTGAGGCG
CTAAATGAAA
CCTTAACGCT
ATGGAACTCG
CCGCCCGACT
GGGCTGGCGA
TGAGCGAAAT
GTAGTGCTTA
CGTTGTCCCG
CATTTGGTAC
AGCGCAGTAA
CCGGCAAAAT
CGCGCCGAAG
GATGTCGCTG
CCGACTGGGC
AATGGAGCGC
CTGCCGGCCC
AGTATCAGCC
CGTCATACTT
GAAGCTAGAC
AGGCTTATCT
TGGACAAGAA
GAAGATCGCT
TGGCCTCGCG
CGCAGATCAG
TTGGAAGAAT
TTGTCCACTA
CGTGAAAGGC
GAGATCACCA
AGGTAGTCGG
CAAATAAAAG
CATTGGTAAC
TGTCAGACCA
AGTTTACTCA
TATATACTTT
AGATTGATTT
AAAACTTCAT
TTTTAATTTA
AAAGGATCTA
GGTGAAGATC
CTTTTTGATA
ATCTCATGAC
CAAAATCCCT
TAACGTGAGT
TTTCGTTCCA
CTGAGCGTCA
GACCCCGTAG
AAAAGATCAA
AGGATCTTCT
TGAGATCCTT
TTTTTCTGCG
CGTAATCTGC
TGCTTGCAAA
CAAAAAAACC
ACCGCTACCA
GCGGTGGTTT
GTTTGCCGGA
TCAAGAGCTA
CCAACTCTTT
TTCCGAAGGT
AACTGGCTTC
AGCAGAGCGC
AGATACCAAA
TACTGTTCTT
CTAGTGTAGC
CGTAGTTAGG
CCACCACTTC
AAGAACTCTG
TAGCACCGCC
TACATACCTC
GCTCTGCTAA
TCCTGTTACC
AGTGGCTGCT
GCCAGTGGCG
ATAAGTCGTG
TCTTACCGGG
TTGGACTCAA
GACGATAGTT
ACCGGATAAG
GCGCAGCGGT
CGGGCTGAAC
GGGGGGTTCG
TGCACACAGC
CCAGCTTGGA
GCGAACGACC
TACACCGAAC
TGAGATACCT
ACAGCGTGAG
CTATGAGAAA
GCGCCACGCT
TCCCGAAGGG
AGAAAGGCGG
ACAGGTATCC
GGTAAGCGGC
AGGGTCGGAA
CAGGAGAGCG
CACGAGGGAG
CTTCCAGGGG
GAAACGCCTG
GTATCTTTAT
AGTCCTGTCG
GGTTTCGCCA
CCTCTGACTT
GAGCGTCGAT
TTTTGTGATG
CTCGTCAGGG
GGGCGGAGCC
TATGGAAAAA
CGCCAGCAAC
GCGGCCTTTT
TACGGTTCCT
GGCCTTTTGC
TGGCCTTTTG
CTCAGGTCTC
ACTCTAGAGG
CATGTGAGGG
CCTATTTCCC
ATGATTCCTT
CATATTTGCA
TATACGATAC
AAGGCTGTTA
GAGAGATAAT
TGGAATTAAT
TTGACTGTAA
ACACAAAGAT
ATTAGTACAA
AATACGTGAC
GTAGAAAGTA
ATAATTTCTT
GGGTAGTTTG
CAGTTTTAAA
ATTATGTTTT
AAAATGGACT
ATCATATGCT
TACCGTAACT
TGAAAGTATT
TCGATTTCTT
GGCTTTATAT
ATCTTGTGGA
AAGGACGAAA
CACCGCATCG
TACGCGTACG
TGTTGTTTTA
GAGCTAGAAA
TAGCAAGTTA
AAATAAGGCT
AGTCCGTTAT
CAACTTGAAA
AAGTGGCACC
GAGTCGGTGC
TTTTTTGTTT
TAGAGCTAGA
AATAGCAAGT
TAAAATAAGG
CTAGTCCGTT
TTTAGCGCGT
GCGCCAATTC
TGCAGACAAA
TGGCTCTA