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?

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

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