QQ:1443274331

010-88891086,80794780

服务电话:

搜索

 北京市昌平区北清路生命科学园北清创意园2-3-102;  Tel:010-88891086,80794780   Fax:010-80794780 
咨询订购QQ: 1443274331   技术支持QQ: 2074750152   邮件订购:bjzblg@126.com      
www.epibio.com.cn  www.epicentre.cn  www.immunoreagents.cn  www.lucigen.com.cn
版权所有 @北京中北林格科技发展有限公司   
京ICP备:13002510号       网站建设:中企动力 北京

进口试剂

>
>
>
CopyControl™ cDNA, Gene & PCR Cloning Kit w/ Electrocomp Cells

进口试剂

CopyControl™ cDNA, Gene & PCR Cloning Kit w/ Electrocomp Cells

浏览量
产品名称:

CopyControl™ cDNA, Gene & PCR Cloning Kit w/ Electrocomp Cells

货号
CCECPCR1
规格
20 Rxns
存储温度
Chemically Competent Cells are stored at -70℃, the remainder is stored at -20℃
没有此类产品
产品描述

Applications

  • Efficient cloning of any insert up to 15 kb, including those that may be unstable or encode proteins that are toxic to the E. coli host cells.

Up to ~25% more cDNA clones can be obtained in a library if the cDNA clones are present in the host cell at only about one copy per cell (P. Carninci and W. Szybalski, personal communication), presumably due to reduced detrimental effects on host cell metabolism and lower toxicity of expressed clone sequences and gene products. The CopyControl™ cDNA, Gene, and PCR Cloning Kits* provide a rapid and improved method (Fig. 1) for cloning any blunt-end DNA fragments (e.g., genomic DNA, PCR products, or cDNA) at single-copy number, with the capability to induce to high-copy numbers as needed (Fig. 2). The CopyControl pCC1™ (Blunt) Vector provided in the kits contains both the E. coli F-factor single-copy origin of replication (oriII) and an inducible high-copy oriV. Replication does not occur using the oriV unless and until the trfA gene in the chromosome of TransforMax™ EPI300™ cells (which is under the control of a tightly regulated promoter) is induced. Thus, CopyControl cDNA, PCR, or gene clones can be grown at single copy to ensure insert stability and successful cloning of sequences that are toxic to the host cell. Then, any desired clone can be induced up to 200 copies per cell for high yields of DNA for all downstream applications.

Benefits

  • Obtain more cDNA or PCR clones and more complete clone libraries (Table 1).
  • Obtain fewer mutated clones. Clones having mutations in genes that are toxic to the host cell are selectively favored at high-copy number.
  • Screen clones for insert size in 1 hour or less without the need for overnight cultures or restriction endonuclease digests (Fig. 3).

 

 Figure 1 (click to enlarge). Efficient, blunt-end cloning of inserts up to 15 kb and screening of clones can be completed in 24 hours using the CopyControl™ cDNA, Gene, and PCR Cloning Kit. Clones can be kept at one copy per cell for clone stability and then, whenever desired, any clone can be induced to high-copy number (up to 200 copies per cell) for high yields of DNA.  

 

Size of PCR Product Cloned cfu/µg of PCR Product Percent Full-Length Clones
1.4 kb >1 x 107 >90%
5 kb 1 x 107 90%
10 kb 3.5 x 106 80%
15 kb 9.2 x 105

30%

 

Table 1. PCR product cloning results with the CopyControl™ System.

Figure 2. Single-copy CopyControl™ clones can be induced to 200 copies per cell. DNA from an equal number of uninduced (–) cells (lane 1) and undiluted induced (+) cells (lane 5) containing a 10-kb CopyControl PCR clone was analyzed by electrophoresis in a 1% agarose gel. Copy number in induced cells was estimated by visually comparing the staining intensity of the diluted induced sample with the uninduced sample. DNA from induced cells was diluted 1:200 (lane 2), 1:100 (lane 3), and 1:50 (lane 4).
Figure 3. Screening of clones from a 5-kb PCR product. The PCR product was cloned using the CopyControl™ cDNA, Gene, and PCR Cloning Kit. Very small portions from nine white colonies were screened using the kit, confirming the presence of the 5-kb insert. Lane M, DNA size ladder; lane 1, 8-kb supercoiled DNA marker; lanes 2-10, randomly chosen clones. Total time for colony screening, including gel electrophoresis, was approximately 60 minutes.  

 

*Covered by issued and/or pending patents.

 

Catalog No. Size

CopyControl™ cDNA, Gene and PCR Cloning Kit (with electrocompetent cells)
   CCECPCR1 20 Reactions
Contents: CopyControl™ pCC1™ (Blunt Cloning-Ready) Vector, PCR Precipitation Solution, PCR 10X Cloning Buffer, End-Repair Enzyme Mix, Fast-Link™ DNA Ligase, EpiLyse™ Solution, EpiBlue™ Solution, CopyControl™ Fosmid Autoinduction Solution, Control PCR Product, Supercoiled DNA Size Marker, Sterile Water, Electrocompetent TransforMax™ EPI300™ E. coli.

CopyControl™ cDNA, Gene and PCR Cloning Kit (with chemically competent cells)
   CCPCR1CC 20 Reactions
Contents: CopyControl™ pCC1™ (Blunt Cloning-Ready) Vector, PCR Precipitation Solution, PCR 10X Cloning Buffer, End-Repair Enzyme Mix, Fast-Link™ DNA Ligase, EpiLyse™ Solution, EpiBlue™ Solution, CopyControl™ Fosmid Autoinduction Solution, Control PCR Product, Supercoiled DNA Size Marker, Sterile Water, Chemically Competent TransforMax™ EPI300™ E. coli.

CopyControl™ Induction Solution
   CCIS125 25 ml
1,000X concentrated solution. Filter sterilized.
 

 

 

未找到相应参数组,请于后台属性模板中添加
暂未实现,敬请期待
暂未实现,敬请期待
上一篇
下一篇