qRT-PCR The reaction mixture for qRT-PCR requires the following: Taq Mix : 1 uL, SYBR Green...
qRT-PCR
The reaction mixture for qRT-PCR requires the following: Taq Mix : 1 uL, SYBR Green Mix: 25 uL, Forward primer : 1 uL, Reverse primer : 1 uL, ROX Reference Dye : 1 uL, Template : 50 ng RNA , Volume up to 50 uL with nuclease free water.
If you are going to make a 20 uL reaction mixture and have a 10 ng/uL RNA sample, how much of each of the components do you need? You also need to say how much water you will use.
Homework Answers
Answer - For 50 microlitre -all the mixture required including Taq mix , SYBR mix, Both forward and reverse primer and ROX dye is total 29 microlitre. And template 50 ng RNA .
That means by dividing 29 from 50 it comes 0.58 .
And it means for making 20 microlitre of reaction mixture by multiplying 0.58 * 20= 11.6 . That means 11.6 uL (microlitre) of reaction mixture is required and rest (8.4 uL ) will be nuclease free water. 200 ng of RNA.
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