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Moleculat ta cloning

Moleculat ta cloning - General Science Questions and Layperson Board

Moleculat ta cloning - General Science Questions and Layperson Board


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Old 03-14-2013, 10:39 AM
Pipette Filler
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Default Moleculat ta cloning



I am working on a virus. trying to clone the 2 genes , one gene i amplifies by using specific primars, but with other gene i am not able to amplfy it . i have tried it at least 10 times, more than that i designed new primar also but didn,t get the band . i want to know were can be the problem.

i tried it with 30 ul reaction volume, 20 ul rxt volume, 50ul rxt volume.

also i used to add tag polymerase at the end

In case of reaction mixture , i use the same constitutents as used with my other gene & also the in same concentration .

Ialso checked it at different annealing temperatures. and at different concentrations of pcr reaction mixture constituents.
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