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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pT7-Cas9(pT7Cas9)
- 库存：
大量
- 供应商：
联迈生物
- 规格：
1ug
质粒名称: pT7-Cas9
其他名称:
质粒大小（bp）: 9036
抗性: Neo/Kan
质粒类型: cloning vector
是否病毒:
启动子:
稳转或瞬转:
组成型或诱导性:
表达水平:
蛋白标签:
测序引物:
GenBank号:
其他属性:
质粒图谱:
质粒序列: AACAAAATATTAACGCTTACAATTTCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCT
TCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACG
ACGTTGTAAAACGACGGCCAGTGCCAAGCTGATCTATACATTGAATCAATATTGGCAATTAGCCATATTAGTCATTGGTT
ATATAGCATAAATCAATATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCA
TGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAG
CCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGA
CGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAA
ACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGC
CTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCA
TGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATT
GACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCA
AATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGAATTTTGTAATACGAC
TCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCCGCCGCGATCGCCATGGATAAGAAA
TACTCAATAGGACTGGATATTGGCACAAATAGCGTCGGATGGGCTGTGATCACTGATGAATATAAGGTTCCTTCTAAAAA
GTTCAAGGTTCTGGGAAATACAGACCGCCACAGTATCAAAAAAAATCTTATAGGGGCTCTTCTGTTTGACAGTGGAGAGA
CAGCCGAAGCTACTAGACTCAAACGGACAGCTAGGAGAAGGTATACAAGACGGAAGAATAGGATTTGTTATCTCCAGGAG
ATTTTTTCAAATGAGATGGCCAAAGTGGATGATAGTTTCTTTCATAGACTTGAAGAGTCTTTTTTGGTGGAAGAAGACAA
GAAGCATGAAAGACATCCTATTTTTGGAAATATAGTGGATGAAGTTGCTTATCACGAGAAATATCCAACTATCTATCATC
TGAGAAAAAAATTGGTGGATTCTACTGATAAAGCCGATTTGCGCCTGATCTATTTGGCCCTGGCCCACATGATTAAGTTT
AGAGGTCATTTTTTGATTGAGGGCGATCTGAATCCTGATAATAGTGATGTGGACAAACTGTTTATCCAGTTGGTGCAAAC
CTACAATCAACTGTTTGAAGAAAACCCTATTAACGCAAGTGGAGTGGATGCTAAAGCCATTCTTTCTGCAAGATTGAGTA
AATCAAGAAGACTGGAAAATCTCATTGCTCAGCTCCCCGGTGAGAAGAAAAATGGCCTGTTTGGGAATCTCATTGCTTTG
TCATTGGGTTTGACCCCTAATTTTAAATCAAATTTTGATTTGGCAGAAGATGCTAAACTCCAGCTTTCAAAAGATACTTA
CGATGATGATCTGGATAATCTGTTGGCTCAAATTGGAGATCAATATGCTGATTTGTTTTTGGCAGCTAAGAATCTGTCAG
ATGCTATTCTGCTTTCAGACATCCTGAGAGTGAATACTGAAATAACTAAGGCTCCCCTGTCAGCTTCAATGATTAAACGC
TACGATGAACATCATCAAGACTTGACTCTTCTGAAAGCCCTGGTTAGACAACAACTTCCAGAAAAGTATAAAGAAATCTT
TTTTGATCAATCAAAAAACGGATATGCAGGTTATATTGATGGCGGCGCAAGCCAAGAAGAATTTTATAAATTTATCAAAC
CAATTCTGGAAAAAATGGATGGTACTGAGGAACTGTTGGTGAAACTGAATAGAGAAGATTTGCTGCGCAAGCAACGGACC
TTTGACAACGGCTCTATTCCCCATCAAATTCACTTGGGTGAGCTGCATGCTATTTTGAGAAGACAAGAAGACTTTTATCC
ATTTCTGAAAGACAATAGAGAGAAGATTGAAAAAATCTTGACTTTTAGGATTCCTTATTATGTTGGTCCATTGGCCAGAG
GCAATAGTAGGTTTGCATGGATGACTCGGAAGTCTGAAGAAACAATTACCCCATGGAATTTTGAAGAAGTTGTCGATAAA
GGTGCTTCAGCTCAATCATTTATTGAACGCATGACAAACTTTGATAAAAATCTTCCAAATGAAAAAGTGCTGCCAAAACA
TAGTTTGCTTTATGAGTATTTTACCGTTTATAACGAATTGACAAAGGTCAAATATGTTACTGAAGGAATGAGAAAACCAG
CATTTCTTTCAGGTGAACAGAAGAAAGCCATTGTTGATCTGCTCTTCAAAACAAATAGGAAAGTGACCGTTAAGCAACTG
AAAGAAGATTATTTCAAAAAAATAGAATGTTTTGATAGTGTTGAAATTTCAGGAGTTGAAGATAGATTTAATGCTTCACT
GGGTACATACCATGATTTGCTGAAAATTATTAAAGATAAAGATTTTTTGGATAATGAAGAAAATGAAGACATCCTGGAGG
ATATTGTTCTGACATTGACCCTGTTTGAAGATAGGGAGATGATTGAGGAAAGACTTAAAACATACGCTCACCTCTTTGAT
GATAAGGTGATGAAACAGCTTAAAAGACGCAGATATACTGGTTGGGGAAGGTTGTCCAGAAAATTGATTAATGGTATTAG
GGATAAGCAATCTGGCAAAACAATACTGGATTTTTTGAAATCAGATGGTTTTGCCAATCGCAATTTTATGCAGCTCATCC
ATGATGATAGTTTGACATTTAAAGAAGACATCCAAAAAGCACAAGTGTCTGGACAAGGCGATAGTCTGCATGAACATATT
GCAAATCTGGCTGGTAGCCCTGCTATTAAAAAAGGTATTCTCCAGACTGTGAAAGTTGTTGATGAATTGGTCAAAGTGAT
GGGGCGGCATAAGCCAGAAAATATCGTTATTGAAATGGCAAGAGAAAATCAGACAACTCAAAAGGGCCAGAAAAATTCCA
GAGAGAGGATGAAAAGAATCGAAGAAGGTATCAAAGAACTGGGAAGTCAGATTCTTAAAGAGCATCCTGTTGAAAATACT
CAATTGCAAAATGAAAAGCTCTATCTCTATTATCTCCAAAATGGAAGAGATATGTATGTGGACCAAGAACTGGATATTAA
TAGGCTGAGTGATTATGATGTCGATCACATTGTTCCACAAAGTTTCCTTAAAGACGATTCAATAGACAATAAGGTCCTGA
CCAGGTCTGATAAAAATAGAGGTAAATCCGATAACGTTCCAAGTGAAGAAGTGGTCAAAAAGATGAAAAACTATTGGAGA
CAACTTCTGAACGCCAAGCTGATCACTCAAAGGAAGTTTGATAATCTGACCAAAGCTGAAAGAGGAGGTTTGAGTGAACT
TGATAAAGCTGGTTTTATCAAACGCCAATTGGTTGAAACTCGCCAAATCACTAAGCATGTGGCACAAATTTTGGATAGTC
GCATGAATACTAAATACGATGAAAATGATAAACTTATTAGAGAGGTTAAAGTGATTACCCTGAAATCTAAACTGGTTTCT
GACTTCAGAAAAGATTTCCAATTCTATAAAGTGAGAGAGATTAACAATTACCATCATGCCCATGATGCCTATCTGAATGC
CGTCGTTGGAACTGCTTTGATTAAGAAATATCCAAAACTTGAAAGCGAGTTTGTCTATGGTGATTATAAAGTTTATGATG
TTAGGAAAATGATTGCTAAGTCTGAGCAAGAAATAGGCAAAGCAACCGCAAAGTATTTCTTTTACTCTAATATCATGAAC
TTCTTCAAAACAGAAATTACACTTGCAAATGGAGAGATTCGCAAACGCCCTCTGATCGAAACTAATGGGGAAACTGGAGA
AATTGTCTGGGATAAAGGGAGAGATTTTGCCACAGTGCGCAAAGTGTTGTCCATGCCCCAAGTCAATATCGTCAAGAAAA
CAGAAGTGCAGACAGGCGGATTCTCTAAGGAGTCAATTCTGCCAAAAAGAAATTCCGACAAGCTGATTGCTAGGAAAAAA
GACTGGGACCCAAAAAAATATGGTGGTTTTGATAGTCCAACCGTGGCTTATTCAGTCCTGGTGGTTGCTAAGGTGGAAAA
AGGGAAATCCAAGAAGCTGAAATCCGTTAAAGAGCTGCTGGGGATCACAATTATGGAAAGAAGTTCCTTTGAAAAAAATC
CCATTGACTTTCTGGAAGCTAAAGGATATAAGGAAGTTAAAAAAGACCTGATCATTAAACTGCCTAAATATAGTCTTTTT
GAGCTGGAAAACGGTAGGAAACGGATGCTGGCTAGTGCCGGAGAACTGCAAAAAGGAAATGAGCTGGCTCTGCCAAGCAA
ATATGTGAATTTTCTGTATCTGGCTAGTCATTATGAAAAGTTGAAGGGTAGTCCAGAAGATAACGAACAAAAACAATTGT
TTGTGGAGCAGCATAAGCATTATCTGGATGAGATTATTGAGCAAATCAGTGAATTTTCTAAGAGAGTTATTCTGGCAGAT
GCCAATCTGGATAAAGTTCTTAGTGCATATAACAAACATAGAGACAAACCAATAAGAGAACAAGCAGAAAATATCATTCA
TCTGTTTACCTTGACCAATCTTGGAGCACCCGCTGCTTTTAAATACTTTGATACAACAATTGATAGGAAAAGATATACCT
CTACAAAAGAAGTTCTGGATGCCACTCTTATCCATCAATCCATCACTGGTCTTTATGAAACACGCATTGATTTGAGTCAG
CTGGGAGGTGACCCCAAGAAAAAACGCAAGGTGGAAGATCCTAAGAAAAAGCGGAAAGTGGACACGCGTACGCGGCCGCT
CGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATTACAAGGATGACGACGATAAGGTTTAAA
CGGCCGGCCGCGGTCATAGCTGTTTCCTGAACAGATCCCGGGTGGCATCCCTGTGACCCCTCCCCAGTGCCTCTCCTGGC
CCTGGAAGTTGCCACTCCAGTGCCCACCAGCCTTGTCCTAATAAAATTAAGTTGCATCATTTTGTCTGACTAGGTGTCCT
TCTATAATATTATGGGGTGGAGGGGGGTGGTATGGAGCAAGGGGCAAGTTGGGAAGACAACCTGTAGGGCCTGCGGGGTC
TATTGGGAACCAAGCTGGAGTGCAGTGGCACAATCTTGGCTCACTGCAATCTCCGCCTCCTGGGTTCAAGCGATTCTCCT
GCCTCAGCCTCCCGAGTTGTTGGGATTCCAGGCATGCATGACCAGGCTCAGCTAATTTTTGTTTTTTTGGTAGAGACGGG
GTTTCACCATATTGGCCAGGCTGGTCTCCAACTCCTAATCTCAGGTGATCTACCCACCTTGGCCTCCCAAATTGCTGGGA
TTACAGGCGTGAACCACTGCTCCCTTCCCTGTCCTTCTGATTTTAAAATAACTATACCAGCAGGAGGACGTCCAGACACA
GCATAGGCTACCTGGCCATGCCCAACCGGTGGGACATTTGAGTTGCTTGCTTGGCACTGTCCTCTCATGCGTTGGGTCCA
CTCAGTAGATGCCTGTTGAATTGGGTACGCGGCCAGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATC
CACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGT
TGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCG
ACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGG
ATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGG
TCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTT
GAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGG
CGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGA
AGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTT
TGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTG
GAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAAT
GAAGTTTTAAATCAATCTAAAGTATATATGAGTAACCTGAGGCTATGGCAGGGCCTGCCGCCCCGACGTTGGCTGCGAGC
CCTGGGCCTTCACCCGAACTTGGGGGGTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTATTGGCCCCAATGGGGTCTCGG
TGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAACCCCGCGTTTATGAACAAACGACCCAACACCGTGCGTTTTATTCT
GTCTTTTTATTGCCGTCATAGCGCGGGTTCCTTCCGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCCCTAGGGTGG
GCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCT
TTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAG
TCCCGCTCAGAAGAACTCGTCAAGAAGGCGATAGAAGGCGATGCGCTGCGAATCGGGAGCGGCGATACCGTAAAGCACGA
GGAAGCGGTCAGCCCATTCGCCGCCAAGCTCTTCAGCAATATCACGGGTAGCCAACGCTATGTCCTGATAGCGATCCGCC
ACACCCAGCCGGCCACAGTCGATGAATCCAGAAAAGCGGCCATTTTCCACCATGATATTCGGCAAGCAGGCATCGCCATG
GGTCACGACGAGATCCTCGCCGTCGGGCATGCTCGCCTTGAGCCTGGCGAACAGTTCGGCTGGCGCGAGCCCCTGATGCT
CTTCGTCCAGATCATCCTGATCGACAAGACCGGCTTCCATCCGAGTACGTGCTCGCTCGATGCGATGTTTCGCTTGGTGG
TCGAATGGGCAGGTAGCCGGATCAAGCGTATGCAGCCGCCGCATTGCATCAGCCATGATGGATACTTTCTCGGCAGGAGC
AAGGTGAGATGACAGGAGATCCTGCCCCGGCACTTCGCCCAATAGCAGCCAGTCCCTTCCCGCTTCAGTGACAACGTCGA
GCACAGCTGCGCAAGGAACGCCCGTCGTGGCCAGCCACGATAGCCGCGCTGCCTCGTCTTGCAGTTCATTCAGGGCACCG
GACAGGTCGGTCTTGACAAAAAGAACCGGGCGCCCCTGCGCTGACAGCCGGAACACGGCGGCATCAGAGCAGCCGATTGT
CTGTTGTGCCCAGTCATAGCCGAATAGCCTCTCCACCCAAGCGGCCGGAGAACCTGCGTGCAATCCATCTTGTTCAATCA
TGCGAAACGATCCTCATCCTGTCTCTTGATCGATCTTTGCAAAAGCCTAGGCCTCCAAAAAAGCCTCCTCACTACTTCTG
GAATAGCTCAGAGGCCGAGGCGGCCTCGGCCTCTGCATAAATAAAAAAAATTAGTCAGCCATGGGGCGGAGAATGGGCGG
AACTGGGCGGAGTTAGGGGCGGGATGGGCGGAGTTAGGGGCGGGACTATGGTTGCTGACTAATTGAGATGCATGCTTTGC
ATACTTCTGCCTGCTGGGGAGCCTGGGGACTTTCCACACCTGGTTGCTGACTAATTGAGATGCATGCTTTGCATACTTCT
GCCTGCTGGGGAGCCTGGGGACTTTCCACACCCTAACTGACACACATTCCACAGCTGGTTCTTTCCGCCTCAGGACTCTT
CCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA
AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGT
GTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCT
CGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACC
TCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACG
TTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGA
TTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTT
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文献和实验CRISPR/Cas9 是一种强大的基因组编辑工具，今天我们介绍 CRISPR/Cas9 系统操作。以下操作说明是基于作者冰糖个人实验室操作，仅供参考。 质粒介绍我们使用的是 Feng Zhang 实验室的系统的 pX330 质粒，如下图所示： 酶切系统 37 ℃, 3 hr；Run 1% gel，回收。Elute 到 50 mL H2O。我们利用 BbsI 消化载体形成粘末端，以利于下游 oligo 退火产物的连接；酶切时请充分，且绝对不能加 CIP 处理
Reaction Mix: It is again convenient to make up a master mix to be aliquotted out to amplify all samples, if they are to use the same primers. If not, modify master mix by simply leaving out primers. Total volume: 50ul / rxn x N + 5-10
选育新方法。而 CRISPR-Cas9 技术的出现，更是极大推动了微生物基因组编辑准确性和效率的提高。【背景】泓迅科技利用 CRISPR-Cas9 技术编辑酵母菌中的一个 920bp 的 ADE1 基因。ADE1 是腺嘌呤合成相关基因，如果 ADE1 失活，则细胞将会积累红色色素，呈现红色菌落。【方法】1、质粒构建2、sgRNA 活性检测3、挑选活性较高的 sgRNA 进行基因组定点编辑4、利用基因功能变化、表性变化和测序等方法检测基因定点修饰结果。【结论】1、从颜色表型变化得知，酵母菌落变成红色菌落，可知 ADE
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