PCR Re-amplification

Klaus.Matthaei at anu.edu.au Klaus.Matthaei at anu.edu.au
Tue Jun 2 22:45:26 EST 1992


In article <1992Jun2.053705.14020 at talon.ucs.orst.edu>
listona at bionette.cgrb.orst.edu (Aaron Liston) writes:
>I would like to hear from anyone who is successfully re-amplifying
>PCR products.  How do you do it?
>Thanks.

Products can be easily re-amplified by using a small amount of the product
(~1-5%) and adding it into a fresh rection mix.  However this I feel is
detrimental since all products including your non-specific junk will
amplify.  It is far better to take this same small amount of product and
amplify in a fresh reaction with nested primers (primers that are inside
the original priming sites and specific to your target).  This should only
amplify your specific target.  In my hands I can amplify less than 100
target molecules in a huge excess of nonspecific DNA.  Feel free to contact
me if you need more information.  Good Luck

Klaus Matthaei
The John Curtin School of Medical Research
The Australian National University
3.6.92




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