Smeared PCR products

Your Name Here you at somehost.somedomain
Sat Feb 25 09:10:02 EST 1995


In article <3idj6b$2pd at ccshst05.cs.uoguelph.ca>, cjerome at uoguelph.ca (Cheryl C Jerome) says:
>
>Hello.  I am wondering if anyone can help me.  I have been having PCR 
>problems for a couple of months.  I have been having consistent smearing 
>with my PCR amplified product.  A bright smear starts at my PCR product 
>and extends to my primers.  Since my PCR product is large (2.8 KB) it is 
>a very large smear which extends to my PCR band.  The smearing is also 
>the negative control.  I have replaced my Taq, my primers, dNTPs, etc... 
>to try to figure out what the problem is.  Also, I am running a positive 
>control with DNA which has worked well in the past.  I had the PCR 
>working with the same conditions and primers for about 2 months.  Then, I 
>was sequencing for about a month and was not running PCR --- when I 
>started to run PCR again, I had the smears and have not been able to 
>resolve the problem.  I have re-autoclaved water, etc....Is it possible 
>there is something in my water source?  what causes smearing? I would 
>appreaciate any suggestions. Thank you. Cheryl

It might be BSA in the TAQ or buffer. Check out methods and reagents in
TIBS Augast 1994.

JIM



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