RT-PCR and gels

Holger Bohnemeier Bohnemeier at ukbf.fu-berlin.de
Tue Aug 1 12:08:35 EST 1995


In article <9507281634.aa00482 at etsuodt.etsu.edu>, betts at ORION.ETSU.EDU (GORDON BETTS) says:
>
>I believe it is possible to run RT then take 3 ul or so and run
>on a 1% agarose gel to verify you have a product. Correct?
>If not, then how can you check to see if your RT worked before 
>wasting reagents on PCR?
>If you can run your RT product on a gel to confirm cDNA is present,
>then could you suggest a procedure that is reliable? Mine's not.
>Gordon Betts, Ph.D.
>Biology Dept.
>East Texas State University
>Commerce, TX 75429-3011
>903/886-5369
>==========================================================================
>A fool and his money deserve to be parted.
>http://www.atlanta.olympic.org/products/bricks.html
>==========================================================================
>
For my part it is a waste of time if you intend to detect your RT-
efficiency by another way than PCR. The interesting cDNA may have
so low concentrations that you are not able to prove its existence
simply by pipetting your post RT sample on a gel.
An alternative might be performing the RT with labeled specific
primers. But talking about saving money I guess you would spend
more in labeling than in performing a PCR.

Bye!



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