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!
* * * * * 1 votes



  • Please log in to reply
No replies to this topic

#1 Dr. Happy

Dr. Happy


  • Members
  • Pip
  • 2 posts

Posted 08 March 2012 - 08:35 AM

hey there
i'm trying to isolate the HVR of HCV-E2..i've extracted RNA from patients' sera using biozol followed by RT-PCR and then first PCR followed by a second PCR using nested primers..i use to have good results before but i failed to clone my fragment..
these days i've purchased a new cloning kit and tried to amplify my fragment as a fresh start and i followed the exact protocol i've optimized over years but i got no results!
to be honest the biozol i'm using is expired but well maintained at +4C and gave me good results just two months ago...it seems O.K and i even i measured it's pH and found it about 4.4...
i changed every single thing starting the 0.2 PCR tubes, RT-enzyme, water, dNTPs and the Taq polymerase i use for first and second PCR and all i've got is primer dimer sometimes with a smear..
what in god's name is going wrong??? RNA extraction?? or my PCR components???!!
if anyone can help me i'll really appreciate it..
best regards for u all

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

©1999-2013 Protocol Online, All rights reserved.