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.


PCR contamination

PCR contamination - PCR - Polymerase Chain Reaction Forum

PCR contamination - 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 06-29-2008, 09:31 AM
Pipette Filler
Points: 428, Level: 8 Points: 428, Level: 8 Points: 428, Level: 8
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2008
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default PCR contamination



I have a problem during PCR program. add templete DNA, dNTP, primer,

polymerase, etc..to ep tube. then I forgot to add oiling U-U

so.. I am nervous for fear of contamination.

Does it matter of this situation? If you shoud be clear up this problem, I will be rearlly preciative of your comment.^_^
Reply With Quote
  #2  
Old 07-02-2008, 01:06 PM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: PCR contamination

Hello Doohe,

welcome to the forum.

Its ok Doohe I am sure you should be ok. Always try things thats how everyone learns. Let us know how the PCR goes and post a gel picture and we can take a look and help.

When you say you forgot to add oiling U-U do you mean you forgot to add oil to the PCR machine?

Reply With Quote
  #3  
Old 08-05-2008, 04:38 PM
Pipette Filler
Points: 731, Level: 14 Points: 731, Level: 14 Points: 731, Level: 14
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2008
Posts: 14
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: PCR contamination

There shouldn't be a problem with contamination. I think that the oil is more to prevent the liquid in the PCR tube from evaporating when the solution is being heated. If your PCR machine can set up a heated lid while running, that might also prevent evaporation.
Reply With Quote
  #4  
Old 08-29-2008, 12:24 AM
Pipette Filler
Points: 3,437, Level: 38 Points: 3,437, Level: 38 Points: 3,437, Level: 38
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2008
Posts: 14
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: PCR contamination

Please i need the help in my problem:
i design primer but when i made PCR, i found may be formation to
primer dimer under the louding dye but the band of primer dimer under
the negative control ( without RNA ) was small in size than the band
under the sample ( more thick and may be 2 band more closed ). i
belived fond 2 band in sample because the negative control and sample
contain same band in the below but found band upper it in the sample
( may be more thickness )at the same conditions. i tried at 47, 52,
55oC annealing. all tempreature give same result with diffrence in 47
the band under negative control more faint but at 55 the band under
sample more thick and light.
please, i need to know this difference meanse any thing or the
difference in size between the band under control and sample don't
means any thing and the 2 bands are primer dimer only.
Thanks for you
Reply With Quote
  #5  
Old 08-29-2008, 12:17 PM
Pipette Filler
Points: 290, Level: 5 Points: 290, Level: 5 Points: 290, Level: 5
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2008
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: PCR contamination

Hi,
I have been getting bands in my control (everything except template DNA)..and when that happens, all the other actual samples also give the exact same bands...and it's an all or none pattern that I have been observing...either all the samples show that sharp bright band that the control has,or at other times all of them show up the correct profile and the control is clean...
i have aliquots of all the ingredients that don't last for more than one PCR run so there is no one 'batch' that i can trace the contamination back to...is it possible that the diluted primers are degrading real fast because one pattern i have noticed is that the first run with a particular primer is alwasy good with all the samples and subsequent runs get bad ..but will degraded primers give bands in the control??

Inputs please!

Thanks,
sg30
Reply With Quote
Reply

Tags
contamination , pcr


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
optimum PCR cycle number and contamination stephmc PCR - Polymerase Chain Reaction Forum 1 10-21-2011 09:51 AM
Contamination of 7H11 agar plates martin rao Microbiology Forum 4 10-31-2010 08:39 PM
PCR contamination makes me mad !! valou PCR - Polymerase Chain Reaction Forum 6 12-04-2008 03:52 PM
qRT-PCR and DNA contamination Jeremy Coate Protocols and Methods Forum 0 05-23-2008 09:55 AM
Methods Digest, Vol 21, Issue 25 Jayanta Tarafdar Protocols and Methods Forum 0 02-23-2007 05:46 PM


All times are GMT. The time now is 02:25 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.13618 seconds with 16 queries