genebridges K001說明書
快速可靠地克隆大DNA分子
在選擇的位置進(jìn)行的DNA修飾
獨(dú)立于限制網(wǎng)站
沒有尺寸限制
簡(jiǎn)單的BAC修飾，如插入選擇標(biāo)記，刪除帶有選擇標(biāo)記基因的片段，插入功能片段和選擇標(biāo)記基因
該試劑盒也可用于細(xì)菌染色體修飾
該試劑盒旨在通過革命性的Red / ET重組技術(shù)對(duì)BAC進(jìn)行修飾 - 這是一種強(qiáng)大的工具，可在2周內(nèi)將修飾（如插入和缺失）引入任何類型的BAC。
紅/ ET重組使用體內(nèi)重組和校對(duì)活動(dòng)來小化構(gòu)建體中不需要的突變。套件中包含了修改BAC所需的所有必要內(nèi)容以及執(zhí)行控制實(shí)驗(yàn)所需的所有材料，旨在讓您熟悉這種新技術(shù)。另外，轉(zhuǎn)化任何大腸桿菌的載體提供了用于應(yīng)用Red / ET重組的感受態(tài)菌株的菌株。pSC101-BAD-gbaAtet帶有四環(huán)素抗性盒。該盒含有溫度敏感的復(fù)制起點(diǎn)（ori），只能在30°C繁殖 - 在37°C時(shí)質(zhì)粒會(huì)丟失。這允許在不再需要質(zhì)粒時(shí)簡(jiǎn)單地去除質(zhì)粒。為了引入突變（即選擇盒，如該試劑盒中包含的新霉素抗性盒），首先必須將同源序列引入選擇盒。這可以通過簡(jiǎn)單的PCR反應(yīng)來完成。這些短的同源序列（每個(gè)50nt）可以根據(jù)您的實(shí)驗(yàn)需要自由選擇。只需選擇要修飾的位置，并使用Red / ET重組技術(shù)將選擇盒直接插入所需的核苷酸。然后需要用誘導(dǎo)的Red / ET重組系統(tǒng)在細(xì)菌中電穿孔PCR產(chǎn)物。在成功進(jìn)行Red / ET重組后，可以使用常規(guī)DNA微量制備方法回收修飾的DNA。
pSC101-BAD-gbaA（tet）是基于Red / ET表達(dá)質(zhì)粒的pSC101 ori（溫度敏感起源）的衍生物
攜帶pSC101-BAD-gbaA（tet）的BAC宿主
用于修飾150kb BAC的陽(yáng)性對(duì)照實(shí)驗(yàn)（攜帶pSC101-BAD-gbaA的宿主中的BAC克隆，用于在BAC骨架中缺失的新PCR產(chǎn)物，在宿主中作為陽(yáng)性重組體的BAC-neo克隆）
方案，質(zhì)粒描述，寡核苷酸圖譜和序列
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
| 貨號(hào) | 品名 | 規(guī)格 | 價(jià)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 |
