PCR yield, can assess by absorbance without electroforesis? - (Jul/06/2008 )

Iam studying about the expression level of FGF-2. I have been done RNA extraction with result in absorbance ratio 260/280 is 1,9. I continue with make first strand cDNA and follow PCR. If PCR process has finished, can I ensure the amplification is success with absorbance assay of DNA in 260nm?because I think if I have known the amplification doesn't success, I should not continue with electroforesis. reversely, If I know the amplification success, I should continue with electroforesis to ensure size of amplificated DNA.How is your opinion?


-fiat29-

bad idea. will not work because in a finished pcr test tube you have a complex mixture of dNTPs, primers, your amplicon, maybe unspecific products, protein (polymerase!) which will all show absorption.

so go straightly to the electrophoresis. there you can see the result with your own eyes.