名称: | pET-28b-RfxCas13d-His |
---|---|
别称: | Plasmid#141322 |
基因长: | |
质粒宿主: | 大肠杆菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF,CRISPR |
片段物种: | |
原核抗性: | Kan |
筛选标记: | |
荧光标记: | |
启动子: | T7 |
复制子: | pBR322 |
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。
CCTGATGCGG
TATTTTCTCC
TTACGCATCT
GTGCGGTATT
TCACACCGCA
ATGGTGCACT
CTCAGTACAA
TCTGCTCTGA
TGCCGCATAG
TTAAGCCAGT
ATACACTCCG
CTATCGCTAC
GTGACTGGGT
CATGGCTGCG
CCCCGACACC
CGCCAACACC
CGCTGACGCG
CCCTGACGGG
CTTGTCTGCT
CCCGGCATCC
GCTTACAGAC
AAGCTGTGAC
CGTCTCCGGG
AGCTGCATGT
GTCAGAGGTT
TTCACCGTCA
TCACCGAAAC
GCGCGAGGCA
GCTGCGGTAA
AGCTCATCAG
CGTGGTCGTG
AAGCGATTCA
CAGATGTCTG
CCTGTTCATC
CGCGTCCAGC
TCGTTGAGTT
TCTCCAGAAG
CGTTAATGTC
TGGCTTCTGA
TAAAGCGGGC
CATGTTAAGG
GCGGTTTTTT
CCTGTTTGGT
CACTGATGCC
TCCGTGTAAG
GGGGATTTCT
GTTCATGGGG
GTAATGATAC
CGATGAAACG
AGAGAGGATG
CTCACGATAC
GGGTTACTGA
TGATGAACAT
GCCCGGTTAC
TGGAACGTTG
TGAGGGTAAA
CAACTGGCGG
TATGGATGCG
GCGGGACCAG
AGAAAAATCA
CTCAGGGTCA
ATGCCAGCGC
TTCGTTAATA
CAGATGTAGG
TGTTCCACAG
GGTAGCCAGC
AGCATCCTGC
GATGCAGATC
CGGAACATAA
TGGTGCAGGG
CGCTGACTTC
CGCGTTTCCA
GACTTTACGA
AACACGGAAA
CCGAAGACCA
TTCATGTTGT
TGCTCAGGTC
GCAGACGTTT
TGCAGCAGCA
GTCGCTTCAC
GTTCGCTCGC
GTATCGGTGA
TTCATTCTGC
TAACCAGTAA
GGCAACCCCG
CCAGCCTAGC
CGGGTCCTCA
ACGACAGGAG
CACGATCATG
CGCACCCGTG
GGGCCGCCAT
GCCGGCGATA
ATGGCCTGCT
TCTCGCCGAA
ACGTTTGGTG
GCGGGACCAG
TGACGAAGGC
TTGAGCGAGG
GCGTGCAAGA
TTCCGAATAC
CGCAAGCGAC
AGGCCGATCA
TCGTCGCGCT
CCAGCGAAAG
CGGTCCTCGC
CGAAAATGAC
CCAGAGCGCT
GCCGGCACCT
GTCCTACGAG
TTGCATGATA
AAGAAGACAG
TCATAAGTGC
GGCGACGATA
GTCATGCCCC
GCGCCCACCG
GAAGGAGCTG
ACTGGGTTGA
AGGCTCTCAA
GGGCATCGGT
CGAGATCCCG
GTGCCTAATG
AGTGAGCTAA
CTTACATTAA
TTGCGTTGCG
CTCACTGCCC
GCTTTCCAGT
CGGGAAACCT
GTCGTGCCAG
CTGCATTAAT
GAATCGGCCA
ACGCGCGGGG
AGAGGCGGTT
TGCGTATTGG
GCGCCAGGGT
GGTTTTTCTT
TTCACCAGTG
AGACGGGCAA
CAGCTGATTG
CCCTTCACCG
CCTGGCCCTG
AGAGAGTTGC
AGCAAGCGGT
CCACGCTGGT
TTGCCCCAGC
AGGCGAAAAT
CCTGTTTGAT
GGTGGTTAAC
GGCGGGATAT
AACATGAGCT
GTCTTCGGTA
TCGTCGTATC
CCACTACCGA
GATATCCGCA
CCAACGCGCA
GCCCGGACTC
GGTAATGGCG
CGCATTGCGC
CCAGCGCCAT
CTGATCGTTG
GCAACCAGCA
TCGCAGTGGG
AACGATGCCC
TCATTCAGCA
TTTGCATGGT
TTGTTGAAAA
CCGGACATGG
CACTCCAGTC
GCCTTCCCGT
TCCGCTATCG
GCTGAATTTG
ATTGCGAGTG
AGATATTTAT
GCCAGCCAGC
CAGACGCAGA
CGCGCCGAGA
CAGAACTTAA
TGGGCCCGCT
AACAGCGCGA
TTTGCTGGTG
ACCCAATGCG
ACCAGATGCT
CCACGCCCAG
TCGCGTACCG
TCTTCATGGG
AGAAAATAAT
ACTGTTGATG
GGTGTCTGGT
CAGAGACATC
AAGAAATAAC
GCCGGAACAT
TAGTGCAGGC
AGCTTCCACA
GCAATGGCAT
CCTGGTCATC
CAGCGGATAG
TTAATGATCA
GCCCACTGAC
GCGTTGCGCG
AGAAGATTGT
GCACCGCCGC
TTTACAGGCT
TCGACGCCGC
TTCGTTCTAC
CATCGACACC
ACCACGCTGG
CACCCAGTTG
ATCGGCGCGA
GATTTAATCG
CCGCGACAAT
TTGCGACGGC
GCGTGCAGGG
CCAGACTGGA
GGTGGCAACG
CCAATCAGCA
ACGACTGTTT
GCCCGCCAGT
TGTTGTGCCA
CGCGGTTGGG
AATGTAATTC
AGCTCCGCCA
TCGCCGCTTC
CACTTTTTCC
CGCGTTTTCG
CAGAAACGTG
GCTGGCCTGG
TTCACCACGC
GGGAAACGGT
CTGATAAGAG
ACACCGGCAT
ACTCTGCGAC
ATCGTATAAC
GTTACTGGTT
TCACATTCAC
CACCCTGAAT
TGACTCTCTT
CCGGGCGCTA
TCATGCCATA
CCGCGAAAGG
TTTTGCGCCA
TTCGATGGTG
TCCGGGATCT
CGACGCTCTC
CCTTATGCGA
CTCCTGCATT
AGGAAGCAGC
CCAGTAGTAG
GTTGAGGCCG
TTGAGCACCG
CCGCCGCAAG
GAATGGTGCA
TGCAAGGAGA
TGGCGCCCAA
CAGTCCCCCG
GCCACGGGGC
CTGCCACCAT
ACCCACGCCG
AAACAAGCGC
TCATGAGCCC
GAAGTGGCGA
GCCCGATCTT
CCCCATCGGT
GATGTCGGCG
ATATAGGCGC
CAGCAACCGC
ACCTGTGGCG
CCGGTGATGC
CGGCCACGAT
GCGTCCGGCG
TAGAGGATCG
AGATCTCGAT
CCCGCGAAAT
TAATACGACT
CACTATAGGG
GAATTGTGAG
CGGATAACAA
TTCCCCTCTA
GAAATAATTT
TGTTTAACTT
TAAGAAGGAG
ATATACCATG
GCGAGCGAGG
CCAGCATCGA
AAAAAAAAAG
TCCTTCGCCA
AGGGCATGGG
CGTGAAGTCC
ACACTCGTGT
CCGGCTCCAA
AGTGTACATG
ACAACCTTCG
CCGAAGGCAG
CGACGCCAGG
CTGGAAAAGA
TCGTGGAGGG
CGACAGCATC
AGGAGCGTGA
ATGAGGGCGA
GGCCTTCAGC
GCTGAAATGG
CCGATAAAAA
CGCCGGCTAT
AAGATCGGCA
ACGCCAAATT
CAGCCATCCT
AAGGGCTACG
CCGTGGTGGC
TAACAACCCT
CTGTATACAG
GACCCGTCCA
GCAGGATATG
CTCGGCCTGA
AGGAAACTCT
GGAAAAGAGG
TACTTCGGCG
AGAGCGCTGA
TGGCAATGAC
AATATTTGTA
TCCAGGTGAT
CCATAACATC
CTGGACATTG
AAAAAATCCT
CGCCGAATAC
ATTACCAACG
CCGCCTACGC
CGTCAACAAT
ATCTCCGGCC
TGGATAAGGA
CATTATTGGA
TTCGGCAAGT
TCTCCACAGT
GTATACCTAC
GACGAATTCA
AAGACCCCGA
GCACCATAGG
GCCGCTTTCA
ACAATAACGA
TAAGCTCATC
AACGCCATCA
AGGCCCAGTA
TGACGAGTTC
GACAACTTCC
TCGATAACCC
CAGACTCGGC
TATTTCGGCC
AGGCCTTTTT
CAGCAAGGAG
GGCAGAAATT
ACATCATCAA
TTACGGCAAC
GAATGCTATG
ACATTCTGGC
CCTCCTGAGC
GGACTGAGGC
ACTGGGTGGT
CCATAACAAC
GAAGAAGAGT
CCAGGATCTC
CAGGACCTGG
CTCTACAACC
TCGATAAGAA
CCTCGACAAC
GAATACATCT
CCACCCTCAA
CTACCTCTAC
GACAGGATCA
CCAATGAGCT
GACCAACTCC
TTCTCCAAGA
ACTCCGCCGC
CAACGTGAAC
TATATTGCCG
AAACTCTGGG
AATCAACCCT
GCCGAATTCG
CCGAACAATA
TTTCAGATTC
AGCATTATGA
AAGAGCAGAA
AAACCTCGGA
TTCAATATCA
CCAAGCTCAG
GGAAGTGATG
CTGGACAGGA
AGGATATGTC
CGAGATCAGG
AAAAATCATA
AGGTGTTCGA
CTCCATCAGG
ACCAAGGTCT
ACACCATGAT
GGACTTTGTG
ATTTATAGGT
ATTACATCGA
AGAGGATGCC
AAGGTGGCTG
CCGCCAATAA
GTCCCTCCCC
GATAATGAGA
AGTCCCTGAG
CGAGAAGGAT
ATCTTTGTGA
TTAACCTGAG
GGGCTCCTTC
AACGACGACC
AGAAGGATGC
CCTCTACTAC
GATGAAGCTA
ATAGAATTTG
GAGAAAGCTC
GAAAATATCA
TGCACAACAT
CAAGGAATTT
AGGGGAAACA
AGACAAGAGA
GTATAAGAAG
AAGGACGCCC
CTAGACTGCC
CAGAATCCTG
CCCGCTGGCC
GTGATGTTTC
CGCCTTCAGC
AAACTCATGT
ATGCCCTGAC
CATGTTCCTG
GATGGCAAGG
AGATCAACGA
CCTCCTGACC
ACCCTGATTA
ATAAATTCGA
TAACATCCAG
AGCTTCCTGA
AGGTGATGCC
TCTCATCGGA
GTCAACGCTA
AGTTCGTGGA
GGAATACGCC
TTTTTCAAAG
ACTCCGCCAA
GATCGCCGAT
GAGCTGAGGC
TGATCAAGTC
CTTCGCTAGA
ATGGGAGAAC
CTATTGCCGA
TGCCAGGAGG
GCCATGTATA
TCGACGCCAT
CCGTATTTTA
GGAACCAACC
TGTCCTATGA
TGAGCTCAAG
GCCCTCGCCG
ACACCTTTTC
CCTGGACGAG
AACGGAAACA
AGCTCAAGAA
AGGCAAGCAC
GGCATGAGAA
ATTTCATTAT
TAATAACGTG
ATCAGCAATA
AAAGGTTCCA
CTACCTGATC
AGATACGGTG
ATCCTGCCCA
CCTCCATGAG
ATCGCCAAAA
ACGAGGCCGT
GGTGAAGTTC
GTGCTCGGCA
GGATCGCTGA
CATCCAGAAA
AAACAGGGCC
AGAACGGCAA
GAACCAGATC
GACAGGTACT
ACGAAACTTG
TATCGGAAAG
GATAAGGGCA
AGAGCGTGAG
CGAAAAGGTG
GACGCTCTCA
CAAAGATCAT
CACCGGAATG
AACTACGACC
AATTCGACAA
GAAAAGGAGC
GTCATTGAGG
ACACCGGCAG
GGAAAACGCC
GAGAGGGAGA
AGTTTAAAAA
GATCATCAGC
CTGTACCTCA
CCGTGATCTA
CCACATCCTC
AAGAATATTG
TCAATATCAA
CGCCAGGTAC
GTCATCGGAT
TCCATTGCGT
CGAGCGTGAT
GCTCAACTGT
ACAAGGAGAA
AGGCTACGAC
ATCAATCTCA
AGAAACTGGA
AGAGAAGGGA
TTCAGCTCCG
TCACCAAGCT
CTGCGCTGGC
ATTGATGAAA
CTGCCCCCGA
TAAGAGAAAG
GACGTGGAAA
AGGAGATGGC
TGAAAGAGCC
AAGGAGAGCA
TTGACAGCCT
CGAGAGCGCC
AACCCCAAGC
TGTATGCCAA
TTACATCAAA
TACAGCGACG
AGAAGAAAGC
CGAGGAGTTC
ACCAGGCAGA
TTAACAGGGA
GAAGGCCAAA
ACCGCCCTGA
ACGCCTACCT
GAGGAACACC
AAGTGGAATG
TGATCATCAG
GGAGGACCTC
CTGAGAATTG
ACAACAAGAC
ATGTACCCTG
TTCAGAAACA
AGGCCGTCCA
CCTGGAAGTG
GCCAGGTATG
TCCACGCCTA
TATCAACGAC
ATTGCCGAGG
TCAATTCCTA
CTTCCAACTG
TACCATTACA
TCATGCAGAG
AATTATCATG
AATGAGAGGT
ACGAGAAAAG
CAGCGGAAAG
GTGTCCGAGT
ACTTCGACGC
TGTGAATGAC
GAGAAGAAGT
ACAACGATAG
GCTCCTGAAA
CTGCTGTGTG
TGCCTTTCGG
CTACTGTATC
CCCAGGTTTA
AGAACCTGAG
CATCGAGGCC
CTGTTCGATA
GGAACGAGGC
CGCCAAGTTC
GACAAGGAGA
AAAAGAAGGT
GTCCGGCAAT
TCCGGATCCG
GAGCGGCCGC
ACTCGAGCAC
CACCACCACC
ACCACTGAGA
TCCGGCTGCT
AACAAAGCCC
GAAAGGAAGC
TGAGTTGGCT
GCTGCCACCG
CTGAGCAATA
ACTAGCATAA
CCCCTTGGGG
CCTCTAAACG
GGTCTTGAGG
GGTTTTTTGC
TGAAAGGAGG
AACTATATCC
GGATTGGCGA
ATGGGACGCG
CCCTGTAGCG
GCGCATTAAG
CGCGGCGGGT
GTGGTGGTTA
CGCGCAGCGT
GACCGCTACA
CTTGCCAGCG
CCCTAGCGCC
CGCTCCTTTC
GCTTTCTTCC
CTTCCTTTCT
CGCCACGTTC
GCCGGCTTTC
CCCGTCAAGC
TCTAAATCGG
GGGCTCCCTT
TAGGGTTCCG
ATTTAGTGCT
TTACGGCACC
TCGACCCCAA
AAAACTTGAT
TAGGGTGATG
GTTCACGTAG
TGGGCCATCG
CCCTGATAGA
CGGTTTTTCG
CCCTTTGACG
TTGGAGTCCA
CGTTCTTTAA
TAGTGGACTC
TTGTTCCAAA
CTGGAACAAC
ACTCAACCCT
ATCTCGGTCT
ATTCTTTTGA
TTTATAAGGG
ATTTTGCCGA
TTTCGGCCTA
TTGGTTAAAA
AATGAGCTGA
TTTAACAAAA
ATTTAACGCG
AATTTTAACA
AAATATTAAC
GCTTACAATT
TAGGTGGCAC
TTTTCGGGGA
AATGTGCGCG
GAACCCCTAT
TTGTTTATTT
TTCTAAATAC
ATTCAAATAT
GTATCCGCTC
ATGAATTAAT
TCTTAGAAAA
ACTCATCGAG
CATCAAATGA
AACTGCAATT
TATTCATATC
AGGATTATCA
ATACCATATT
TTTGAAAAAG
CCGTTTCTGT
AATGAAGGAG
AAAACTCACC
GAGGCAGTTC
CATAGGATGG
CAAGATCCTG
GTATCGGTCT
GCGATTCCGA
CTCGTCCAAC
ATCAATACAA
CCTATTAATT
TCCCCTCGTC
AAAAATAAGG
TTATCAAGTG
AGAAATCACC
ATGAGTGACG
ACTGAATCCG
GTGAGAATGG
CAAAAGTTTA
TGCATTTCTT
TCCAGACTTG
TTCAACAGGC
CAGCCATTAC
GCTCGTCATC
AAAATCACTC
GCATCAACCA
AACCGTTATT
CATTCGTGAT
TGCGCCTGAG
CGAGACGAAA
TACGCGATCG
CTGTTAAAAG
GACAATTACA
AACAGGAATC
GAATGCAACC
GGCGCAGGAA
CACTGCCAGC
GCATCAACAA
TATTTTCACC
TGAATCAGGA
TATTCTTCTA
ATACCTGGAA
TGCTGTTTTC
CCGGGGATCG
CAGTGGTGAG
TAACCATGCA
TCATCAGGAG
TACGGATAAA
ATGCTTGATG
GTCGGAAGAG
GCATAAATTC
CGTCAGCCAG
TTTAGTCTGA
CCATCTCATC
TGTAACATCA
TTGGCAACGC
TACCTTTGCC
ATGTTTCAGA
AACAACTCTG
GCGCATCGGG
CTTCCCATAC
AATCGATAGA
TTGTCGCACC
TGATTGCCCG
ACATTATCGC
GAGCCCATTT
ATACCCATAT
AAATCAGCAT
CCATGTTGGA
ATTTAATCGC
GGCCTAGAGC
AAGACGTTTC
CCGTTGAATA
TGGCTCATAA
CACCCCTTGT
ATTACTGTTT
ATGTAAGCAG
ACAGTTTTAT
TGTTCATGAC
CAAAATCCCT
TAACGTGAGT
TTTCGTTCCA
CTGAGCGTCA
GACCCCGTAG
AAAAGATCAA
AGGATCTTCT
TGAGATCCTT
TTTTTCTGCG
CGTAATCTGC
TGCTTGCAAA
CAAAAAAACC
ACCGCTACCA
GCGGTGGTTT
GTTTGCCGGA
TCAAGAGCTA
CCAACTCTTT
TTCCGAAGGT
AACTGGCTTC
AGCAGAGCGC
AGATACCAAA
TACTGTCCTT
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
CTCACATGTT
CTTTCCTGCG
TTATCCCCTG
ATTCTGTGGA
TAACCGTATT
ACCGCCTTTG
AGTGAGCTGA
TACCGCTCGC
CGCAGCCGAA
CGACCGAGCG
CAGCGAGTCA
GTGAGCGAGG
AAGCGGAAGA
GCG