Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > PCR - Polymerase Chain Reaction Forum
Register Search Today's Posts Mark Forums Read

PCR - Polymerase Chain Reaction Forum PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


Sequencing troubles of a PCR product

Sequencing troubles of a PCR product - PCR - Polymerase Chain Reaction Forum

Sequencing troubles of a PCR product - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 09-19-2008, 10:42 AM
Pipette Filler
Points: 357, Level: 7 Points: 357, Level: 7 Points: 357, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2008
Location: Portugal
Posts: 10
Thanks: 0
Thanked 0 Times in 0 Posts
Default Sequencing troubles of a PCR product



Hello,
maybe this is not the rigth place to post this question but i didint know where to post it...
i am with troubles in sequencing a PCR product; this is the image I get of my sequencing:

when the sequencing that i want is over it seems like there is another sequencing, but i am sure that this is not a contamination because i have diferent samples...

the point is: when i do the blast on ncbi i get the blast of my sequencing, i dont know why but it does another blast (I got 2 blasts) of the second sequencing i am relly relly confused... my lab partners don't know wath is happening as well...
could anyone hellp me? do you know of any article or site that i could consult?
thanks anyway..
Reply With Quote
  #2  
Old 09-19-2008, 03:11 PM
Graduate Student
Points: 2,923, Level: 35 Points: 2,923, Level: 35 Points: 2,923, Level: 35
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2006
Location: Ithaca, NY
Posts: 145
Thanks: 0
Thanked 17 Times in 12 Posts
Default Re: Sequencing troubles of a PCR product

I'm not quite sure what the problem is. It looks like you dropped off the end of your PCR product. You might have some tiny amount of something else in there (perhaps template) but it's not significantly interfering with your primary peaks. Trim the sequence at the end of your good read (about 260 bases) and blast just what you want to blast.

Or am I missing the point of your question?
Reply With Quote
  #3  
Old 09-23-2008, 03:06 PM
Pipette Filler
Points: 357, Level: 7 Points: 357, Level: 7 Points: 357, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2008
Location: Portugal
Posts: 10
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Sequencing troubles of a PCR product

thanks very much for the response Kmunson779! You get the poin of my question and you had hellp me in fact! I will do what you said :-)
Reply With Quote
Reply

Tags
pcr , product , sequencing , troubles


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
PCR Product Sequencing moleculardude PCR - Polymerase Chain Reaction Forum 1 06-09-2009 12:53 PM
PCR product sequencing majd Molecular Biology Techniques 1 12-19-2008 05:57 PM
PCR product sequencing majd PCR - Polymerase Chain Reaction Forum 0 08-04-2008 09:23 AM
Sequencing - Low Product Melody Protocols and Methods Forum 1 09-02-2005 11:14 AM


All times are GMT. The time now is 12:29 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 0.12396 seconds with 16 queries