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

>
>
>
>
Direct Lysis Plasmid96 DNA Purification Kit

进口试剂

Direct Lysis Plasmid96 DNA Purification Kit

浏览量
产品名称:

Direct Lysis Plasmid96 DNA Purification Kit

货号
PLS84596
规格
5 x 96 well preps
存储温度
Room Temperature
没有此类产品
产品描述
Applications
  • Rapid, direct-lysis preparation of plasmid DNA for sequencing in a 96-well format.

The Direct Lysis Plasmid96 Kit isolates plasmid DNA by direct lysis of a 200-µl overnight culture, eliminating the need for centrifugation of the culture and resuspension of the cell pellet (Fig. 1). The kit is designed for rapid preparation of sequencing templates in a 96-well format from either high-copy or single-copy plasmids sized up to 40 kb (Figs. 2 and 3). The kit uses a modified alkaline lysis procedure that incorporates a proprietary RNase Blend to remove the RNA efficiently. The typical yield from a 200-µl overnight culture of a high-copy plasmid vector is sufficient for up to 50 end-sequencing reactions. A copy-number-induced 40-kb plasmid will yield DNA for six to eight sequencing reactions.

The kit includes reagents and plastics for high-throughput DNA extraction, and is suitable for robotic automation procedures.

Benefits

  • No cell harvesting or resuspension of the cell pellet.
  • Inoculation of culture in a 200-µl volume eliminates the need for 2-ml, deep-well growth plates.
  • Reduces the amount of media used to under 20 ml per plate compared to 150 ml with a standard 1.2-ml prep.
  • Low final DNA suspension volume (50 µl).
  • Protocol complete in as little as 25 minutes.
  • Perform ~50 end-sequencing reactions from a high-copy plasmid, with average read length of 700-900 bases (Fig. 3).
  • Low-profile filter-receiver plate block (H57 x L127 x W85 mm).
Overnight culture:
Add lysis buffer; mix.
Add neutralization buffer; mix.
Figure 1
No cell harvesting.
No use of deep-well
culture plate.
Arrow

Clear the lysate:
Vacuum filtration or
centrifugation, 1 min.

  Arrow
Precipitate the DNA:
Centrifuge, 10 min.
Wash with 70% EtOH, 5 min.
Resuspend the DNA.
Figure 1. Overview of the Direct Lysis Plasmid96 purification process.
 

 

Figure 2. Examination of the quality of plasmid DNA. DNA was isolated from 200-µl overnight cultures of randomly picked clones from a shotgun library (2-6 kb) in a 96-well plate using the Direct Lysis Plasmid96 Kit. A 5-µl aliquot of each 50-µl prep was analyzed on a 1% agarose gel in 1X TAE buffer for 2 hours. Lane M, Supercoiled DNA Ladder; lanes 1-18, supercoiled plasmid DNA. Figure 3. Sequence analysis confirms plasmid DNA quality. Sequencing was performed using 2 µl of DNA isolated from a plasmid and M13 Forward primer, and was analyzed on an ABI® 3730 Analyzer in the UWBC DNA Sequencing and Synthesis Facility (Madison, WI).

 

Catalog No. Size

Direct Lysis Plasmid96 DNA Purification Kit
PLS84596 5 x 96-well preps
Contents:
Five culture plates, five 96-well fritted filter plates, five collection plates, plate sealers, gas-permeable sealers, and reagents for five 96-well preps.

 

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