genebridges K001说明书
Quick & Easy BAC Modification Kit
快速简便的BAC修改套件
- ID: K001
特征
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快速可靠地克隆大DNA分子
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在选择的位置进行的DNA修饰
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独立于限制网站
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没有尺寸限制
应用
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简单的BAC修饰，如插入选择标记，删除带有选择标记基因的片段，插入功能片段和选择标记基因
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该试剂盒也可用于细菌染色体修饰
描述
该试剂盒旨在通过革命性的Red / ET重组技术对BAC进行修饰 – 这是一种强大的工具，可在2周内将修饰（如插入和缺失）引入任何类型的BAC。
红/ ET重组使用体内重组和校对活动来小化构建体中不需要的突变。套件中包含了修改BAC所需的所有必要内容以及执行控制实验所需的所有材料，旨在让您熟悉这种新技术。另外，转化任何大肠杆菌的载体提供了用于应用Red / ET重组的感受态菌株的菌株。pSC101-BAD-gbaAtet带有四环素抗性盒。该盒含有温度敏感的复制起点（ori），只能在30°C繁殖 – 在37°C时质粒会丢失。这允许在不再需要质粒时简单地去除质粒。为了引入突变（即选择盒，如该试剂盒中包含的新霉素抗性盒），首先必须将同源序列引入选择盒。这可以通过简单的PCR反应来完成。这些短的同源序列（每个50nt）可以根据您的实验需要自由选择。只需选择要修饰的位置，并使用Red / ET重组技术将选择盒直接插入所需的核苷酸。然后需要用诱导的Red / ET重组系统在细菌中电穿孔PCR产物。在成功进行Red / ET重组后，可以使用常规DNA微量制备方法回收修饰的DNA。
内容
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pSC101-BAD-gbaA（tet）是基于Red / ET表达质粒的pSC101 ori（温度敏感起源）的衍生物
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携带pSC101-BAD-gbaA（tet）的BAC宿主
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用于修饰150kb BAC的阳性对照实验（携带pSC101-BAD-gbaA的宿主中的BAC克隆，用于在BAC骨架中缺失的新PCR产物，在宿主中作为阳性重组体的BAC-neo克隆）
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方案，质粒描述，寡核苷酸图谱和序列
Sequences
Tn5-kanR/neoR selection cassette
Tn5-promoter kanR/neoR
TGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGA AGCCCTGCAAAGTAAACTGGATGGCTTTCTTGCCGCCAAGGATCTGATGGCGCAG GGGATCAAGATCTGATCAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAA GATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATG ACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGC GCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAA CTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCG CAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGA AGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCC ATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCAT TCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGG TCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAA CTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCC ATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATT CATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCT ACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGC TTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGA
CGAGTTCTTCTGACTCGAG
| 货号 | 品名 | 规格 | 价格 |
| K001 | * Quick & Easy BAC Modification Kit | Kit | 13900 |
| K002 | * Counter Selection BAC Modification Kit | Kit | 16400 |
| K003 | * BAC Subcloning Kit | Kit | 13900 |
| K004 | * Quick & Easy Conditional Knock Out Kit – FRT | Kit | 13900 |
| K005 | * Quick & Easy Conditional Knock Out Kit – loxP | Kit | 13900 |
| K006 | *Quick & Easy E.Coli Gene Deletion Kit | Kit | 13900 |
| K008 | *Direct-cloning-proficient E.coli strain GB05-dir | Kit | 12400 |
| K009 | *Recombineering-proficient E.coli strain GB08-red | Kit | 12400 |
| A001 | Pro- and Eukaryotic Neomycin Selection Cassette (PGK-gb2-neo) | 50ng/ | 5980 |
| A002 | FRT flanked, Pro and Euk. Neomycin Select. Cassette (FRT-PGK-gb2-neo-FRT) | 50ng/ | 5980 |
| A003 | loxP flanked, Pro and Euk. Neomycin Select. Cassette (loxP-PGK-gb2-neo-loxP) | 50ng/ | 5980 |
| A004 | FRT flanked, Pro and Euk. Neomycin Select. Cassette plus loxP site(FRT-PGK-gb2-neo-FRT-loxP) | 50ng/ | 5980 |
| A005 | FRT flanked, Pro and Euk. Neomycin Select. Cassette plus loxP site 2nd version (loxP-FRT-PGK-gb2-neo-FRT) | 50ng/ | 5980 |
| A006 | FRT flanked Chloramphenicol Select. Cassette (FRT-cm-FRT) | 50ng/ | 5980 |
| A007 | loxP flanked Chloramphenicol Select. Cassette (loxP-cm-loxP) | 50ng/ | 5980 |
| A008 | FRT flanked Ampicillin Select. Cassette (FRT-amp-FRT) | 50ng/ | 5980 |
| A009 | loxP flanked Ampicillin Select. Cassette (loxP-amp-loxP) | 50ng/ | 5980 |
| A010 | FRT flanked, Pro and Euk. Hygromycin Select. Cassette (FRT-PGK-gb2-hygro-FRT) | 50ng/ | 5980 |
| A011 | loxP flanked, Pro and Euk. Hygromycin Select. Cassette (loxP-PGK-gb2-hygro-loxP) | 50ng/ | 5980 |
| A012 | iCre-FRT-neo-FRT (codon improved Cre(iCre) with attached FRT flanked, Pro- and Eukaryotic Neomycin selection cassette | 50ng/ | 6500 |
| A013 | iCreERT2-FRT-neo-FRT (ligand-inducible iCre with attached FRT flanked, Pro- and Eukaryotic Neomycin Selection cassette) | 50ng/ | 6500 |
| A104 | Enhanced Flp Expression Plasmid:707-FLPe; tet | 0.2ug/ | 5980 |
| A105 | Enhanced Flp Expression Plasmid:708-FLPe;cm | 0.2ug/ | 5980 |
| A106 | Enhanced Flp Expression Plasmid:709-FLPe;amp | 0.2ug/ | 5980 |
| A107 | Enhanced Flp Expression Plasmid:710-FLPe;km | 0.2ug/ | 5980 |
| A112 | Cre Expression Plasmid:705-Cre;cm | 0.2ug/ | 5980 |
| A113 | Cre Expression Plasmid:706-Cre;tet | 0.2ug/ | 5980 |
| A114 | Cre Expression Plasmid:704-Cre;amp | 0.2ug/ | 5980 |
| A201 | Enhanced Eukaryotic FLP Expression Plasmid: pCAGGS-FLPe (Academia) | 0.2ug/ | 7500 |
| A203 | Eukaryotic FLPo Recombinase Expression Plasmid: pCAGGS-FLPo (Academia)* | 0.2ug/ | 7500 |
| A204 | Eukaryotic Cre Recombinase Expression Plasmid: pCAGGS-Cre (Academia)* | 0.2ug/ | 7500 |
| A205 | Eukaryotic Der Recombinase Expression Plasmid: CAGGS-Dre(Academia)* | 0.2ug/ | 7500 |
| A301 | Arabinose-inducible Prokaryotic Cre Recombinase Expression Plasmid: pSC101-BAD-Cre | 0.2ug/ | 5980 |
| A302 | Arabinose-inducible Prokaryotic Dre Recombinase Expression Plasmid: pSC101-BAD-Dre* | 0.2ug/ | 5980 |
| A303 | Arabinose-inducible Prokaryotic FLPe Recombinase Expression Plasmid: pSC101-BAD-FLPe (tet) | 0.2ug/ | 5980 |