re-amp of PCR -- problems

[rsorg at chmeds.ac.nz]

In article <1995Jan11.121642.1 at csc.canterbury.ac.nz>, zool201 at csc.canterbury.
ac.nz writes:
> To whom it may concern
> 
> I'm trying to reamplify a very low concentrartion of primary PCR
> product and am ending up with smears every time.
> I don't want ot re-purify the primary product because of the very low 
> concentration.
> 
> Any hints would be greatfully accepted.
> 
> Andrew Holyoake 
> PhD student
> University of Canterbury
> Christchurch 
> New Zealand

Dear Andrew,

I think you have a problem which is quite common and which I had to face too
when I was trying to reamplify a PCR product. Two things might help:

1. Dilute your primary PCR product at least 1:10,000 and try it again.

or

2. Run your PCR product on an agarose gel, cut out the band, add 1 ml of 
   water and boil it for 5min. I actually could amplify all my primary PCR 
   products by doing this and using 1-5ul for the secondary PCR.


-----------------------------------
Ruediger V. Sorg, Ph.D.            |
Christchurch School of Medicine    |
Haematology/Immunology Research    |
Christchurch, New Zealand          |
E-MAIL RSORG at CHMEDS.AC.NZ          |
PHONE #64 (3) 364-0553             |
FAX   #64 (3) 364-0750             |