Multi primers PCR

Karel Wernars lwlkarel at rivm.nl
Thu Dec 8 07:42:49 EST 1994


In Article <1994Dec6.161419.1 at leif>, mmai at kean.ucs.mun.ca wrote:
>
>Hi. Does anyone have the experience to do multi primers PCR? It means that
>more than one gene fragments are amplified with two or more pairs of primers.
>If it works this technique will be very good the check low abundance mRNA
>with a control PCR band. Thanx in advance.

I have done PCR-detection of enterohaemorrhagic E.coli in enrichment-cultures
from cattle feces. For this two sets of primers were used to amplify fragments
of two virulence related genes (eae and slt-II). Doing this as a four-primer
assay was no problem with respect to specificity. However, sensitivity of detection
decreased because the total amount of amplified DNA was equal compared to
the two-primer assay. Therefor it is my advise to stick to the usual two-primer
assay if sensitivity is an important aspect of your experiments.


Karel Wernars, The Netherlands



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