PCR to the PCR!! - (Apr/02/2005 )
i heard today about a strange thing (at least strange for me )
someone told me that u can Make a better quality your PCR product by using a previously made PCR product as a tempate instead of the original idea of using DNA as template
is that true !!!!!!!
Yes it's true...enjoy it
you can certainly use a PCR product as a template.
In my experience, it gives more DNA product.
regarding quality, well, you can't get anything out of a PCR product as template except the PCR product itself (did I confuse everyone there?).
no you were clear
i was just woundring , say u have a PCR product that was non specific (with a smear appearing instead of typical bands , would you epect much from using that PCR product as a template ??
and wht isf that prodct has primer dimer instead of smearing could it also give better results as a template ?????
Hmm, in that case, I don't think it will help you at all. Because now you have all these PCR products that your primer will be able to bind to. On top of that, your template is still in there, swimming with your smear or primer-dimer.
I would try to work out the optimal conditions for the original PCR.
Might I add, this is the whole idea behind a nested PCR.
Primers designed internal to the original set are used in the second round to "enhance" the template yield.
You can perform a second round of PCR with the original primer set as you all have pointed out.
you are right about the cooncept of nested PCR ,,,it didnt cross my mind
anyhow i well use only good quality PCR products as templates to get
excellent PCR products (hopefully)