Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques
Register Search Today's Posts Mark Forums Read

Molecular Biology Techniques Molecular Biology Forum. Includes forums for common molecular biology techniques.


Ampliying part of a virus genome

Ampliying part of a virus genome - Molecular Biology Techniques

Ampliying part of a virus genome - Molecular Biology Forum. Includes forums for common molecular biology techniques.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 02-08-2010, 09:19 PM
Pipette Filler
Points: 14, Level: 1 Points: 14, Level: 1 Points: 14, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2010
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Ampliying part of a virus genome



Hi all,
I'm trying to make a standard curve for my Real-time PCR. I need to amplify part of my virus genome which is a ss RNA, positive sense genome. I need to use genomic template by boiling my virus and use its genome as my template, PCR a part of it and ligate it ino a vector from TOPO cloning kit. My question is that whether I need to make a cDNA from the template first and then use it in the PCR reaction or there is another way to perform this process. I've never made such a thing and need some help. Please drop me a line if you have any idea.
Thanks a lot.
Reply With Quote
  #2  
Old 02-10-2010, 07:01 PM
danfive's Avatar
Nobel Laureate
Points: 8,612, Level: 64 Points: 8,612, Level: 64 Points: 8,612, Level: 64
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Location: Houston TX
Posts: 969
Thanks: 12
Thanked 184 Times in 154 Posts
Default Re: Ampliying part of a virus genome

Yes.

Perform Reverse Transcription using just the Reverse Primer.

Take cDNA and PCR amplify (using Fwd and Rev primers, lower number of cycles than usual).

From there make several dilutions of PCR product and use as vector for ligation reaction.

Transform the e.coli as directed by TOPO protocol.
Reply With Quote
Reply

Tags
ampliying , genome , part , virus


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
Human Cytome Project - an idea - Update 19 April 2005 Peter Van Osta Cell Biology and Cell Culture 1 06-01-2009 02:17 PM
A Human Cytome Project - an idea - Update 14 March 2005 Peter Van Osta Cell Biology and Cell Culture 0 03-14-2005 02:27 PM
Human Cytome Project - Update 6 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 0 01-06-2005 11:18 AM


All times are GMT. The time now is 09:03 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16835 seconds with 16 queries