名称: | pCMV-T7-SpRY-P2A-EGFP (RTW4830) |
---|---|
别称: | Plasmid #139989 |
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: | Amp |
筛选标记: | |
荧光标记: | 绿色 |
启动子: | CMV |
复制子: | pUC |
拷贝数: | |
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。
CGGCAAACAA
ACCACCGCTG
GTAGCGGTGG
TTTTTTTGTT
TGCAAGCAGC
AGATTACGCG
CAGAAAAAAA
GGATCTCAAG
AAGATCCTTT
GATCTTTTCT
ACGGGGTCTG
ACGCTCAGTG
GAACGAAAAC
TCACGTTAAG
GGATTTTGGT
CATGAGATTA
TCAAAAAGGA
TCTTCACCTA
GATCCTTTTA
AATTAAAAAT
GAAGTTTTAA
ATCAATCTAA
AGTATATATG
AGTAAACTTG
GTCTGACAGT
TACCAATGCT
TAATCAGTGA
GGCACCTATC
TCAGCGATCT
GTCTATTTCG
TTCATCCATA
GTTGCCTGAC
TCCCCGTCGT
GTAGATAACT
ACGATACGGG
AGGGCTTACC
ATCTGGCCCC
AGTGCTGCAA
TGATACCGCG
AGACCCACGC
TCACCGGCTC
CAGATTTATC
AGCAATAAAC
CAGCCAGCCG
GAAGGGCCGA
GCGCAGAAGT
GGTCCTGCAA
CTTTATCCGC
CTCCATCCAG
TCTATTAATT
GTTGCCGGGA
AGCTAGAGTA
AGTAGTTCGC
CAGTTAATAG
TTTGCGCAAC
GTTGTTGCCA
TTGCTACAGG
CATCGTGGTG
TCACGCTCGT
CGTTTGGTAT
GGCTTCATTC
AGCTCCGGTT
CCCAACGATC
AAGGCGAGTT
ACATGATCCC
CCATGTTGTG
CAAAAAAGCG
GTTAGCTCCT
TCGGTCCTCC
GATCGTTGTC
AGAAGTAAGT
TGGCCGCAGT
GTTATCACTC
ATGGTTATGG
CAGCACTGCA
TAATTCTCTT
ACTGTCATGC
CATCCGTAAG
ATGCTTTTCT
GTGACTGGTG
AGTACTCAAC
CAAGTCATTC
TGAGAATAGT
GTATGCGGCG
ACCGAGTTGC
TCTTGCCCGG
CGTCAATACG
GGATAATACC
GCGCCACATA
GCAGAACTTT
AAAAGTGCTC
ATCATTGGAA
AACGTTCTTC
GGGGCGAAAA
CTCTCAAGGA
TCTTACCGCT
GTTGAGATCC
AGTTCGATGT
AACCCACTCG
TGCACCCAAC
TGATCTTCAG
CATCTTTTAC
TTTCACCAGC
GTTTCTGGGT
GAGCAAAAAC
AGGAAGGCAA
AATGCCGCAA
AAAAGGGAAT
AAGGGCGACA
CGGAAATGTT
GAATACTCAT
ACTCTTCCTT
TTTCAATATT
ATTGAAGCAT
TTATCAGGGT
TATTGTCTCA
TGAGCGGATA
CATATTTGAA
TGTATTTAGA
AAAATAAACA
AATAGGGGTT
CCGCGCACAT
TTCCCCGAAA
AGTGCCACCT
GACGTCGACG
GATCGGGAGA
TCGATCTCCC
GATCCCCTAG
GGTCGACTCT
CAGTACAATC
TGCTCTGATG
CCGCATAGTT
AAGCCAGTAT
CTGCTCCCTG
CTTGTGTGTT
GGAGGTCGCT
GAGTAGTGCG
CGAGCAAAAT
TTAAGCTACA
ACAAGGCAAG
GCTTGACCGA
CAATTGCATG
AAGAATCTGC
TTAGGGTTAG
GCGTTTTGCG
CTGCTTCGCG
ATGTACGGGC
CAGATATACG
CGTTGACATT
GATTATTGAC
TAGTTATTAA
TAGTAATCAA
TTACGGGGTC
ATTAGTTCAT
AGCCCATATA
TGGAGTTCCG
CGTTACATAA
CTTACGGTAA
ATGGCCCGCC
TGGCTGACCG
CCCAACGACC
CCCGCCCATT
GACGTCAATA
ATGACGTATG
TTCCCATAGT
AACGCCAATA
GGGACTTTCC
ATTGACGTCA
ATGGGTGGAG
TATTTACGGT
AAACTGCCCA
CTTGGCAGTA
CATCAAGTGT
ATCATATGCC
AAGTACGCCC
CCTATTGACG
TCAATGACGG
TAAATGGCCC
GCCTGGCATT
ATGCCCAGTA
CATGACCTTA
TGGGACTTTC
CTACTTGGCA
GTACATCTAC
GTATTAGTCA
TCGCTATTAC
CATGGTGATG
CGGTTTTGGC
AGTACATCAA
TGGGCGTGGA
TAGCGGTTTG
ACTCACGGGG
ATTTCCAAGT
CTCCACCCCA
TTGACGTCAA
TGGGAGTTTG
TTTTGGCACC
AAAATCAACG
GGACTTTCCA
AAATGTCGTA
ACAACTCCGC
CCCATTGACG
CAAATGGGCG
GTAGGCGTGT
ACGGTGGGAG
GTCTATATAA
GCAGAGCTGG
TTTAGTGAAC
CGTCAGATCC
GCTAGAGATC
CGCGGCCGCT
AATACGACTC
ACTATAGGGA
GAGCCGCCAC
CATGGACAAG
AAGTACAGCA
TCGGCCTGGA
CATCGGCACC
AACTCTGTGG
GCTGGGCCGT
GATCACCGAC
GAGTACAAGG
TGCCCAGCAA
GAAATTCAAG
GTGCTGGGCA
ACACCGACCG
GCACAGCATC
AAGAAGAACC
TGATCGGAGC
CCTGCTGTTC
GACAGCGGCG
AAACAGCCGA
GAGAACCCGG
CTGAAGAGAA
CCGCCAGAAG
AAGATACACC
AGACGGAAGA
ACCGGATCTG
CTATCTGCAA
GAGATCTTCA
GCAACGAGAT
GGCCAAGGTG
GACGACAGCT
TCTTCCACAG
ACTGGAAGAG
TCCTTCCTGG
TGGAAGAGGA
TAAGAAGCAC
GAGCGGCACC
CCATCTTCGG
CAACATCGTG
GACGAGGTGG
CCTACCACGA
GAAGTACCCC
ACCATCTACC
ACCTGAGAAA
GAAACTGGTG
GACAGCACCG
ACAAGGCCGA
CCTGCGGCTG
ATCTATCTGG
CCCTGGCCCA
CATGATCAAG
TTCCGGGGCC
ACTTCCTGAT
CGAGGGCGAC
CTGAACCCCG
ACAACAGCGA
CGTGGACAAG
CTGTTCATCC
AGCTGGTGCA
GACCTACAAC
CAGCTGTTCG
AGGAAAACCC
CATCAACGCC
AGCGGCGTGG
ACGCCAAGGC
CATCCTGTCT
GCCAGACTGA
GCAAGAGCAG
ACGGCTGGAA
AATCTGATCG
CCCAGCTGCC
CGGCGAGAAG
AAGAATGGCC
TGTTCGGAAA
CCTGATTGCC
CTGAGCCTGG
GCCTGACCCC
CAACTTCAAG
AGCAACTTCG
ACCTGGCCGA
GGATGCCAAA
CTGCAGCTGA
GCAAGGACAC
CTACGACGAC
GACCTGGACA
ACCTGCTGGC
CCAGATCGGC
GACCAGTACG
CCGACCTGTT
TCTGGCCGCC
AAGAACCTGT
CCGACGCCAT
CCTGCTGAGC
GACATCCTGA
GAGTGAACAC
CGAGATCACC
AAGGCCCCCC
TGAGCGCCTC
TATGATCAAG
AGATACGACG
AGCACCACCA
GGACCTGACC
CTGCTGAAAG
CTCTCGTGCG
GCAGCAGCTG
CCTGAGAAGT
ACAAAGAGAT
TTTCTTCGAC
CAGAGCAAGA
ACGGCTACGC
CGGCTACATT
GACGGCGGAG
CCAGCCAGGA
AGAGTTCTAC
AAGTTCATCA
AGCCCATCCT
GGAAAAGATG
GACGGCACCG
AGGAACTGCT
CGTGAAGCTG
AACAGAGAGG
ACCTGCTGCG
GAAGCAGCGG
ACCTTCGACA
ACGGCAGCAT
CCCCCACCAG
ATCCACCTGG
GAGAGCTGCA
CGCCATTCTG
CGGCGGCAGG
AAGATTTTTA
CCCATTCCTG
AAGGACAACC
GGGAAAAGAT
CGAGAAGATC
CTGACCTTCC
GCATCCCCTA
CTACGTGGGC
CCTCTGGCCA
GGGGAAACAG
CAGATTCGCC
TGGATGACCA
GAAAGAGCGA
GGAAACCATC
ACCCCCTGGA
ACTTCGAGGA
AGTGGTGGAC
AAGGGCGCTT
CCGCCCAGAG
CTTCATCGAG
CGGATGACCA
ACTTCGATAA
GAACCTGCCC
AACGAGAAGG
TGCTGCCCAA
GCACAGCCTG
CTGTACGAGT
ACTTCACCGT
GTATAACGAG
CTGACCAAAG
TGAAATACGT
GACCGAGGGA
ATGAGAAAGC
CCGCCTTCCT
GAGCGGCGAG
CAGAAAAAGG
CCATCGTGGA
CCTGCTGTTC
AAGACCAACC
GGAAAGTGAC
CGTGAAGCAG
CTGAAAGAGG
ACTACTTCAA
GAAAATCGAG
TGCTTCGACT
CCGTGGAAAT
CTCCGGCGTG
GAAGATCGGT
TCAACGCCTC
CCTGGGCACA
TACCACGATC
TGCTGAAAAT
TATCAAGGAC
AAGGACTTCC
TGGACAATGA
GGAAAACGAG
GACATTCTGG
AAGATATCGT
GCTGACCCTG
ACACTGTTTG
AGGACAGAGA
GATGATCGAG
GAACGGCTGA
AAACCTATGC
CCACCTGTTC
GACGACAAAG
TGATGAAGCA
GCTGAAGCGG
CGGAGATACA
CCGGCTGGGG
CAGGCTGAGC
CGGAAGCTGA
TCAACGGCAT
CCGGGACAAG
CAGTCCGGCA
AGACAATCCT
GGATTTCCTG
AAGTCCGACG
GCTTCGCCAA
CAGAAACTTC
ATGCAGCTGA
TCCACGACGA
CAGCCTGACC
TTTAAAGAGG
ACATCCAGAA
AGCCCAGGTG
TCCGGCCAGG
GCGATAGCCT
GCACGAGCAC
ATTGCCAATC
TGGCCGGCAG
CCCCGCCATT
AAGAAGGGCA
TCCTGCAGAC
AGTGAAGGTG
GTGGACGAGC
TCGTGAAAGT
GATGGGCCGG
CACAAGCCCG
AGAACATCGT
GATCGAAATG
GCCAGAGAGA
ACCAGACCAC
CCAGAAGGGA
CAGAAGAACA
GCCGCGAGAG
AATGAAGCGG
ATCGAAGAGG
GCATCAAAGA
GCTGGGCAGC
CAGATCCTGA
AAGAACACCC
CGTGGAAAAC
ACCCAGCTGC
AGAACGAGAA
GCTGTACCTG
TACTACCTGC
AGAATGGGCG
GGATATGTAC
GTGGACCAGG
AACTGGACAT
CAACCGGCTG
TCCGACTACG
ATGTGGACCA
TATCGTGCCT
CAGAGCTTTC
TGAAGGACGA
CTCCATCGAC
AACAAGGTGC
TGACCAGAAG
CGACAAGAAC
CGGGGCAAGA
GCGACAACGT
GCCCTCCGAA
GAGGTCGTGA
AGAAGATGAA
GAACTACTGG
CGGCAGCTGC
TGAACGCCAA
GCTGATTACC
CAGAGAAAGT
TCGACAATCT
GACCAAGGCC
GAGAGAGGCG
GCCTGAGCGA
ACTGGATAAG
GCCGGCTTCA
TCAAGAGACA
GCTGGTGGAA
ACCCGGCAGA
TCACAAAGCA
CGTGGCACAG
ATCCTGGACT
CCCGGATGAA
CACTAAGTAC
GACGAGAATG
ACAAGCTGAT
CCGGGAAGTG
AAAGTGATCA
CCCTGAAGTC
CAAGCTGGTG
TCCGATTTCC
GGAAGGATTT
CCAGTTTTAC
AAAGTGCGCG
AGATCAACAA
CTACCACCAC
GCCCACGACG
CCTACCTGAA
CGCCGTCGTG
GGAACCGCCC
TGATCAAAAA
GTACCCTAAG
CTGGAAAGCG
AGTTCGTGTA
CGGCGACTAC
AAGGTGTACG
ACGTGCGGAA
GATGATCGCC
AAGAGCGAGC
AGGAAATCGG
CAAGGCTACC
GCCAAGTACT
TCTTCTACAG
CAACATCATG
AACTTTTTCA
AGACCGAGAT
TACCCTGGCC
AACGGCGAGA
TCCGGAAGCG
GCCTCTGATC
GAGACAAACG
GCGAAACCGG
GGAGATCGTG
TGGGATAAGG
GCCGGGATTT
TGCCACCGTG
CGGAAAGTGC
TGAGCATGCC
CCAAGTGAAT
ATCGTGAAAA
AGACCGAGGT
GCAGACAGGC
GGCTTCAGCA
AAGAGTCTAT
CAGACCCAAG
AGGAACAGCG
ATAAGCTGAT
CGCCAGAAAG
AAGGACTGGG
ACCCTAAGAA
GTACGGCGGC
TTCCTGTGGC
CCACCGTGGC
CTATTCTGTG
CTGGTGGTGG
CCAAAGTGGA
AAAGGGCAAG
TCCAAGAAAC
TGAAGAGTGT
GAAAGAGCTG
CTGGGGATCA
CCATCATGGA
AAGAAGCAGC
TTCGAGAAGA
ATCCCATCGA
CTTTCTGGAA
GCCAAGGGCT
ACAAAGAAGT
GAAAAAGGAC
CTGATCATCA
AGCTGCCTAA
GTACTCCCTG
TTCGAGCTGG
AAAACGGCCG
GAAGAGAATG
CTGGCCTCTG
CCAAGCAGCT
GCAGAAGGGA
AACGAACTGG
CCCTGCCCTC
CAAATATGTG
AACTTCCTGT
ACCTGGCCAG
CCACTATGAG
AAGCTGAAGG
GCTCCCCCGA
GGATAATGAG
CAGAAACAGC
TGTTTGTGGA
ACAGCACAAG
CACTACCTGG
ACGAGATCAT
CGAGCAGATC
AGCGAGTTCT
CCAAGAGAGT
GATCCTGGCC
GACGCTAATC
TGGACAAAGT
GCTGTCCGCC
TACAACAAGC
ACCGGGATAA
GCCCATCAGA
GAGCAGGCCG
AGAATATCAT
CCACCTGTTT
ACCCTGACCA
GACTGGGAGC
CCCTAGAGCC
TTCAAGTACT
TTGACACCAC
CATCGACCCC
AAGCAGTACA
GAAGCACCAA
AGAGGTGCTG
GACGCCACCC
TGATCCACCA
GAGCATCACC
GGCCTGTACG
AGACACGGAT
CGACCTGTCT
CAGCTGGGAG
GCGACGGATC
CGGCGGAGGC
GGAAGCGGGA
AAAGAACCGC
CGACGGCAGC
GAATTCGAGC
CCAAGAAGAA
GAGGAAAGTC
TCGAGCGGAG
GCGACTACAA
AGACCATGAC
GGTGATTATA
AAGATCATGA
CATCGATTAC
AAGGATGACG
ATGACAAGGG
AGGCGGTGGA
AGCGGCGCAA
CAAACTTCTC
TCTGCTGAAA
CAAGCCGGAG
ATGTCGAAGA
GAATCCTGGA
CCGATGGTGA
GCAAGGGCGA
GGAGCTGTTC
ACCGGGGTGG
TGCCCATCCT
GGTCGAGCTG
GACGGCGACG
TAAACGGCCA
CAAGTTCAGC
GTGTCCGGCG
AGGGCGAGGG
CGATGCCACC
TACGGCAAGC
TGACCCTGAA
GTTCATCTGC
ACCACCGGCA
AGCTGCCCGT
GCCCTGGCCC
ACCCTCGTGA
CCACCCTGAC
CTACGGCGTG
CAGTGCTTCA
GCCGCTACCC
CGACCACATG
AAGCAGCACG
ACTTCTTCAA
GTCCGCCATG
CCCGAAGGCT
ACGTCCAGGA
GCGCACCATC
TTCTTCAAGG
ACGACGGCAA
CTACAAGACC
CGCGCCGAGG
TGAAGTTCGA
GGGCGACACC
CTGGTGAACC
GCATCGAGCT
GAAGGGCATC
GACTTCAAGG
AGGACGGCAA
CATCCTGGGG
CACAAGCTGG
AGTACAACTA
CAACAGCCAC
AACGTCTATA
TCATGGCCGA
CAAGCAGAAG
AACGGCATCA
AGGTGAACTT
CAAGATCCGC
CACAACATCG
AGGACGGCAG
CGTGCAGCTC
GCCGACCACT
ACCAGCAGAA
CACCCCCATC
GGCGACGGCC
CCGTGCTGCT
GCCCGACAAC
CACTACCTGA
GCACCCAGTC
CGCCCTGAGC
AAAGACCCCA
ACGAGAAGCG
CGATCACATG
GTCCTGCTGG
AGTTCGTGAC
CGCCGCCGGG
ATCACTCTCG
GCATGGACGA
GCTGTACAAG
TGACCGGTCA
TCATCACCAT
CACCATTGAG
TTTAAACCCG
CTGATCAGCC
TCGACTGTGC
CTTCTAGTTG
CCAGCCATCT
GTTGTTTGCC
CCTCCCCCGT
GCCTTCCTTG
ACCCTGGAAG
GTGCCACTCC
CACTGTCCTT
TCCTAATAAA
ATGAGGAAAT
TGCATCGCAT
TGTCTGAGTA
GGTGTCATTC
TATTCTGGGG
GGTGGGGTGG
GGCAGGACAG
CAAGGGGGAG
GATTGGGAAG
ACAATAGCAG
GCATGCTGGG
GATGCGGTGG
GCTCTATGGC
TTCTGAGGCG
GAAAGAACCA
GCTGGGGCTC
GATACCGTCG
ACCTCTAGCT
AGAGCTTGGC
GTAATCATGG
TCATAGCTGT
TTCCTGTGTG
AAATTGTTAT
CCGCTCACAA
TTCCACACAA
CATACGAGCC
GGAAGCATAA
AGTGTAAAGC
CTAGGGTGCC
TAATGAGTGA
GCTAACTCAC
ATTAATTGCG
TTGCGCTCAC
TGCCCGCTTT
CCAGTCGGGA
AACCTGTCGT
GCCAGCTGCA
TTAATGAATC
GGCCAACGCG
CGGGGAGAGG
CGGTTTGCGT
ATTGGGCGCT
CTTCCGCTTC
CTCGCTCACT
GACTCGCTGC
GCTCGGTCGT
TCGGCTGCGG
CGAGCGGTAT
CAGCTCACTC
AAAGGCGGTA
ATACGGTTAT
CCACAGAATC
AGGGGATAAC
GCAGGAAAGA
ACATGTGAGC
AAAAGGCCAG
CAAAAGGCCA
GGAACCGTAA
AAAGGCCGCG
TTGCTGGCGT
TTTTCCATAG
GCTCCGCCCC
CCTGACGAGC
ATCACAAAAA
TCGACGCTCA
AGTCAGAGGT
GGCGAAACCC
GACAGGACTA
TAAAGATACC
AGGCGTTTCC
CCCTGGAAGC
TCCCTCGTGC
GCTCTCCTGT
TCCGACCCTG
CCGCTTACCG
GATACCTGTC
CGCCTTTCTC
CCTTCGGGAA
GCGTGGCGCT
TTCTCATAGC
TCACGCTGTA
GGTATCTCAG
TTCGGTGTAG
GTCGTTCGCT
CCAAGCTGGG
CTGTGTGCAC
GAACCCCCCG
TTCAGCCCGA
CCGCTGCGCC
TTATCCGGTA
ACTATCGTCT
TGAGTCCAAC
CCGGTAAGAC
ACGACTTATC
GCCACTGGCA
GCAGCCACTG
GTAACAGGAT
TAGCAGAGCG
AGGTATGTAG
GCGGTGCTAC
AGAGTTCTTG
AAGTGGTGGC
CTAACTACGG
CTACACTAGA
AGAACAGTAT
TTGGTATCTG
CGCTCTGCTG
AAGCCAGTTA
CCTTCGGAAA
AAGAGTTGGT
AGCTCTTGAT
C