Why platinum taq amplification of a gene is giving smear on the gel?

Neha vinod Ahuja @Neha_Ahuja3

07 July 2016 1 6K Report

i am amplifying 6.7kb fragment using HiFi platinum taq. conditions used are:

annealing temperature 72 deg celsius

gDNA 600ng

MgSO4 1.5mM

But i am getting smear again and again. Kindly suggest how to amplify this big fragment.

Disha Mody

Hi Neha! Did u use all the controls? No DNA control, No Taq control.

One of ur reagents could be contaminated or u may be using sheared DNA to which ur primers may not have been able to bind. Using the above control may help u figure out where things may be going wrong..

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