sequencing pcr products

Karen Roberts leehrat at mindspring.com
Thu Apr 6 20:02:15 EST 2000


No one ever sat me down and explained the rationale behind od 260 and
280.
All I know is the conversion factor for ds dna is 50ug/ml and for rna is
40ug/ml.  I have also seen a conversion factor of 20ug/ml; what does
that refer to?

What is the conversion factor to use for quanitating purified pcr
products?  Also, when evaluating the quality of a purified pcr product
(as opposed to say, a "dirty" plamsmid), is there anything that can be
used as an index of quality like the 260/280 ratio?

We are going to use pcr product as our template to sequence.  Is too
much primer in the sequencing reaction a bad thing?

TIA,
Karen





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