Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

Genomic library DIFFICULTIES!

puc19 lipase gene no RE digest stressed

  • Please log in to reply
No replies to this topic




  • Active Members
  • Pip
  • 25 posts

Posted 18 June 2015 - 02:46 AM

Dear researchers please help


Im actual;y working on cold active lipases. Currently trying to look for novel lipases. Here is what i did:


I did intial digest with Sau3A1 with my genomic DNA with different concentration and mange to get a band of 10kb to 2 kb in which the lipase genes are believed to be in.


I ligated my product into puc 19 (vector) and host of DH5 alpha. I seem to get good no of colonies on the plate however when I test with RE digest (double digest) i just get a smearing band. Im using Eco R1 with Hind111 buffer tango


Please help. Attached is the gel with harvested plasmid. All of them seem to have  a band which is above 10 kbIM000530 (new).jpg


Please help!!! Do i change the plasmid, or the RE?

Also tagged with one or more of these keywords: puc19, lipase gene, no RE digest, stressed

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.