名称: | pBLO ddd64.1 |
---|---|
别称: | Plasmid #123121;CasX |
基因长: | |
质粒宿主: | 哺乳细胞 |
质粒用途: | 基因编辑 |
片段类型: | CRISPR,Cas9 |
片段物种: | |
原核抗性: | Chl |
筛选标记: | |
荧光标记: | |
启动子: | Tet |
复制子: | p15A |
拷贝数: | 低 |
感受态: | DH5a |
温度: | 37度 |
背景载体: | |
正向引物: | |
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。
CGACCAGAAC
AAGCTGAAGC
CCGAGATGGA
CGAGAAGGGC
AATCTGACCA
CAGCCGGCTT
TGCCTGCTCT
CAGTGTGGCC
AGCCTCTGTT
CGTGTACAAG
CTGGAACAGG
TGTCCGAGAA
AGGCAAGGCC
TACACCAACT
ACTTCGGCAG
ATGTAACGTG
GCCGAGCACG
AGAAGCTGAT
TCTGCTGGCC
CAGCTGAAAC
CTGAGAAGGA
CTCTGATGAG
GCCGTGACCT
ACAGCCTGGG
CAAGTTTGGA
CAGAGAGCCC
TGGACTTCTA
CAGCATCCAC
GTGACCAAAG
AAAGCACACA
CCCCGTGAAG
CCCCTGGCTC
AGATCGCCGG
CAATAGATAC
GCCTCTGGAC
CTGTGGGCAA
AGCCCTGTCC
GATGCCTGCA
TGGGAACAAT
CGCCAGCTTC
CTGAGCAAGT
ACCAGGACAT
CATCATCGAG
CACCAGAAGG
TGGTCAAGGG
CAACCAGAAG
AGACTGGAAA
GCCTGAGGGA
GCTGGCCGGC
AAAGAGAACC
TGGAATACCC
CAGCGTGACC
CTGCCTCCTC
AGCCTCACAC
AAAAGAAGGC
GTGGACGCCT
ACAACGAAGT
GATCGCCAGA
GTGAGAATGT
GGGTCAACCT
GAACCTGTGG
CAGAAGCTGA
AACTGTCCAG
GGACGACGCC
AAGCCTCTGC
TGAGACTGAA
GGGCTTCCCT
AGCTTCCCCG
TGGTGGAAAG
AAGGGAGAAT
GAGGTGGACT
GGTGGAACAC
CATCAACGAA
GTCAAGAAAC
TGATCGATGC
CAAGAGAGAC
ATGGGCAGAG
TGTTTTGGAG
CGGAGTGACC
GCCGAGAAGA
GAAACACCAT
CCTGGAAGGC
TACAACTACC
TGCCTAACGA
GAACGACCAC
AAGAAGAGAG
AGGGCAGCCT
GGAAAACCCT
AAGAAGCCCG
CCAAGAGACA
GTTCGGCGAC
CTGCTGCTGT
ACCTGGAGAA
GAAGTATGCC
GGCGACTGGG
GCAAAGTGTT
CGATGAGGCC
TGGGAGAGAA
TCGACAAGAA
GATTGCCGGC
CTGACCAGCC
ACATCGAGAG
AGAGGAAGCC
AGAAACGCCG
AGGACGCCCA
GTCTAAAGCC
GTGCTGACAG
ATTGGCTGAG
AGCCAAGGCC
AGCTTCGTGC
TGGAAAGGCT
GAAAGAAATG
GATGAGAAAG
AGTTCTACGC
CTGCGAGATC
CAGCTGCAGA
AGTGGTACGG
GGACCTGAGA
GGCAATCCCT
TTGCCGTGGA
AGCCGAGAAC
AGAGTGGTGG
ACATCAGCGG
CTTTAGCATC
GGCTCTGACG
GCCACTCCAT
CCAGTACAGG
AACCTGCTGG
CCTGGAAGTA
CCTGGAAAAT
GGCAAGAGAG
AGTTCTACCT
GCTGATGAAC
TACGGCAAGA
AGGGCAGAAT
CAGATTCACC
GACGGCACCG
ACATCAAGAA
AAGCGGCAAG
TGGCAGGGCC
TGCTGTATGG
CGGCGGAAAG
GCCAAAGTGA
TCGACCTGAC
CTTCGATCCC
GACGACGAGC
AGCTGATCAT
TCTGCCTCTG
GCCTTCGGCA
CCAGACAGGG
CAGAGAGTTC
ATCTGGAACG
ATCTGCTGTC
CCTGGAAACC
GGCCTGATCA
AGCTGGCCAA
CGGCAGGGTG
ATCGAGAAAA
CCATCTACAA
CAAGAAAATC
GGCAGAGATG
AGCCCGCACT
GTTTGTGGCC
CTGACCTTTG
AGAGGAGAGA
AGTGGTCGAT
CCCAGCAACA
TCAAGCCCGT
GAACCTGATC
GGAGTGGCCA
GAGGCGAGAA
TATCCCTGCC
GTGATTGCCC
TGACAGACCC
TGAGGGATGT
CCCCTGCCTG
AGTTTAAGGA
TAGCTCTGGC
GGCCCTACCG
ACATCCTGAG
AATCGGCGAG
GGCTACAAAG
AGAAGCAGAG
AGCCATCCAG
GCCGCCAAAG
AGGTGGAACA
GAGAAGAGCC
GGCGGATACA
GCAGAAAGTT
CGCCAGCAAG
TCCAGAAACC
TGGCCGATGA
CATGGTCAGG
AACAGCGCCA
GGGACCTGTT
CTACCACGCC
GTGACACATG
ATGCCGTGCT
GGTGTTCGCG
AACCTGTCAA
GAGGCTTCGG
CAGACAGGGA
AAAAGAACCT
TCATGACCGA
GAGACAGTAC
ACGAAGATGG
AAGATTGGCT
CACCGCCAAA
CTGGCCTACG
AGGGCCTGAC
AAGCAAGACC
TACCTGTCCA
AGACACTGGC
CCAGTACACC
TCCAAGACCT
GCTCCAATTG
CGGCTTCACC
ATCACCACCG
CCGACTACGA
TGGAATGCTC
GTGAGACTGA
AGAAAACCTC
CGACGGCTGG
GCCACCACAC
TGAACAACAA
AGAGCTGAAA
GCCGAGGGCC
AGATCACCTA
CTACAACAGA
TACAAGAGGC
AGACCGTCGA
GAAAGAGCTG
TCCGCCGAAC
TCGACAGACT
GAGCGAGGAA
TCCGGCAACA
ACGACATCAG
CAAGTGGACC
AAGGGACGCA
GAGATGAGGC
CCTGTTCCTG
CTGAAGAAGA
GGTTCAGCCA
CAGACCTGTG
CAAGAGCAGT
TTGTGTGCCT
GGATTGCGGC
CACGAAGTGC
ACGCAGCTGA
ACAGGCCGCT
CTGAATATCG
CCAGAAGCTG
GCTGTTCCTG
AACAGCAACA
GCACCGAGTT
CAAGAGCTAC
AAGAGCGGCA
AGCAGCCTTT
CGTTGGAGCC
TGGCAGGCCT
TCTACAAGAG
AAGGCTCAAA
GAAGTGTGGA
AGCCCAACGC
CTGACTCGAG
TAAGGATCTC
CAGGCATCAA
ATAAAACGAA
AGGCTCAGTC
GAAAGACTGG
GCCTTTCGTT
TTATCTGTTG
TTTGTCGGTG
AACGCTCTCT
ACTAGAGTCA
CACTGGCTCA
CCTTCGGGTG
GGCCTTTCTG
CGTTTATACC
TAGGGATATA
TTCCGCTTCC
TCGCTCACTG
ACTCGCTACG
CTCGGTCGTT
CGACTGCGGC
GAGCGGAAAT
GGCTTACGAA
CGGGGCGGAG
ATTTCCTGGA
AGATGCCAGG
AAGATACTTA
ACAGGGAAGT
GAGAGGGCCG
CGGCAAAGCC
GTTTTTCCAT
AGGCTCCGCC
CCCCTGACAA
GCATCACGAA
ATCTGACGCT
CAAATCAGTG
GTGGCGAAAC
CCGACAGGAC
TATAAAGATA
CCAGGCGTTT
CCCCCTGGCG
GCTCCCTCGT
GCGCTCTCCT
GTTCCTGCCT
TTCGGTTTAC
CGGTGTCATT
CCGCTGTTAT
GGCCGCGTTT
GTCTCATTCC
ACGCCTGACA
CTCAGTTCCG
GGTAGGCAGT
TCGCTCCAAG
CTGGACTGTA
TGCACGAACC
CCCCGTTCAG
TCCGACCGCT
GCGCCTTATC
CGGTAACTAT
CGTCTTGAGT
CCAACCCGGA
AAGACATGCA
AAAGCACCAC
TGGCAGCAGC
CACTGGTAAT
TGATTTAGAG
GAGTTAGTCT
TGAAGTCATG
CGCCGGTTAA
GGCTAAACTG
AAAGGACAAG
TTTTGGTGAC
TGCGCTCCTC
CAAGCCAGTT
ACCTCGGTTC
AAAGAGTTGG
TAGCTCAGAG
AACCTTCGAA
AAACCGCCCT
GCAAGGCGGT
TTTTTCGTTT
TCAGAGCAAG
AGATTACGCG
CAGACCAAAA
CGATCTCAAG
AAGATCATCT
TATTAATCAG
ATAAAATATT
TCTAGATTTC
AGTGCAATTT
ATCTCTTCAA
ATGTAGCACC
TGAAGTCAGC
CCCATACGAT
ATAAGTTGTT
ACTAGTGCTT
GGATTCTCAC
CAATAAAAAA
CGCCCGGCGG
CAACCGAGCG
TTCTGAACAA
ATCCAGATGG
AGTTCTGAGG
TCATTACTGG
ATCTATCAAC
AGGAGTCCAA
GCGAGCTCGA
TATCAAATTA
CGCCCCGCCC
TGCCACTCAT
CGCAGTACTG
TTGTAATTCA
TTAAGCATTC
TGCCGACATG
GAAGCCATCA
CAAACGGCAT
GATGAACCTG
AATCGCCAGC
GGCATCAGCA
CCTTGTCGCC
TTGCGTATAA
TATTTGCCCA
TGGTGAAAAC
GGGGGCGAAG
AAGTTGTCCA
TATTGGCCAC
GTTTAAATCA
AAACTGGTGA
AACTCACCCA
GGGATTGGCT
GACACGAAAA
ACATATTCTC
AATAAACCCT
TTAGGGAAAT
AGGCCAGGTT
TTCACCGTAA
CACGCCACAT
CTTGCGAATA
TATGTGTAGA
AACTGCCGGA
AATCGTCGTG
GTATTCACTC
CAGAGCGATG
AAAACGTTTC
AGTTTGCTCA
TGGAAAACGG
TGTAACAAGG
GTGAACACTA
TCCCATATCA
CCAGCTCACC
GTCTTTCATT
GCCATACGAA
ATTCCGGATG
AGCATTCATC
AGGCGGGCAA
GAATGTGAAT
AAAGGCCGGA
TAAAACTTGT
GCTTATTTTT
CTTTACGGTC
TTTAAAAAGG
CCGTAATATC
CAGCTGAACG
GTCTGGTTAT
AGGTACATTG
AGCAACTGAC
TGAAATGCCT
CAAAATGTTC
TTTACGATGC
CATTGGGATA
TATCAACGGT
GGTATATCCA
GTGATTTTTT
TCTCCATTTT
AGCTTCCTTA
GCTCCTGAAA
ATCTCGATAA
CTCAAAAAAT
ACGCCCGGTA
GTGATCTTAT
TTCATTATGG
TGAAAGTTGG
AACCTCTTAC
GTGCCGATCA
ACGTTTCATT
TTCGCCAGAT
ATCGACGTCT
TAAGACCCAC
TTTCACATTT
AAGTTGTTTT
TCTAATCCGC
ATATGATCAA
TTCAAGGCCG
AATAAGAAGG
CTGGCTCTGC
ACCTTGGTGA
TCAAATAATT
CGATAGCTTG
TCGTAATAAT
GGCGGCATAC
TATCAGTAGT
AGGTGTTTCC
CTTTCTTCTT
TAGCGACTTG
ATGCTCTTGA
TCTTCCAATA
CGCAACCTAA
AGTAAAATGC
CCCACAGCGC
TGAGTGCATA
TAATGCATTC
TCTAGTGAAA
AACCTTGTTG
GCATAAAAAG
GCTAATTGAT
TTTCGAGAGT
TTCATACTGT
TTTTCTGTAG
GCCGTGTACC
TAAATGTACT
TTTGCTCCAT
CGCGATGACT
TAGTAAAGCA
CATCTAAAAC
TTTTAGCGTT
ATTACGTAAA
AAATCTTGCC
AGCTTTCCCC
TTCTAAAGGG
CAAAAGTGAG
TATGGTGCCT
ATCTAACATC
TCAATGGCTA
AGGCGTCGAG
CAAAGCCCGC
TTATTTTTTA
CATGCCAATA
CAATGTAGGC
TGCTCTACAC
CTAGCTTCTG
GGCGAGTTTA
CGGGTTGTTA
AACCTTCGAT
TCCGACCTCA
TTAAGCAGCT
CTAATGCGCT
GTTAATCACT
TTACTTTTAT
CTAATCTAGA
CATCATTAAT
TCCTAATTTT
TGTTGACACT
CTATCGTTGA
TAGAGTTATT
TTACCACTCC
CTATCAGTGA
TAGAGAAAAG
AATTCAAAAG
ATCTAAAGAG
GAGAAAGGAT
CTATGGAAAA
GAGAATCAAC
AAGATCAGGA
AGAAGCTGAG
CGCCGACAAC
GCCACCAAGC
CTGTGTCTAG
GAGTGGCCCC
ATGAAAACCC
TGCTCGTCAG
AGTGATGACC
GACGACCTGA
AGAAGAGACT
GGAAAAGAGG
AGGAAGAAAC
CCGAAGTGAT
GCCCCAAGTG
ATCAGCAACA
ACGCCGCCAA
CAACCTGAGA
ATGCTGCTGG
ACGACTACAC
CAAGATGAAG
GAAGCCATCC
TGCAGGTCTA
CTGGCAAGAG
TTCAAGGACG
ACCACGTGGG
CCTGATGTGC
AAGTTTGCCC
AGCCTGCCAG
CAAGAAGAT