Question

BioChemistry

How do I calculate the yield (Amount) of RNA for the following sample?

As well as calculate the volume of each RNA preparation required for the cDNA synthesis

For the following:

Sample A -

610.75 ng/uL

A260/A280: 2.03

A260/A230: 1.65

Answer 1

Ans:- The yield for RNA is very easy to calculate. During your RNA isolation procedure, what is the final volume of RNA that you obtained. Means at last step of RNA isolation either you have dissolved the RNA pellet in some volume of Nuclese free water or using column you have eluted the RNA. Suppose, you have used 20 ul for elution or dissolving.

Now, you measured the concentration of RNA and find that it is 610.75 ng/ul.

So, the total yield of RNA will be 20 * 610.75 ng

That 12,215 ng was total yield. Now, one more things you can do if you know the with how much cells you have strated or used for RNA isolation. Then you can tell 12,215 ng/ cells number.

Now, for cDNA synthesis you really have to look how much RNA is required for synthesis of cDNA. Most of the cDNA synthesis kit mentioned use 2 to 5 ug of RNA. Now, suppose we use 5 ug of RNA then we have to add 8.2 ul of your mentioned RNA concentration.

Now, once you have 5 ug of RNA in tube see the catalogue of cDNA synthesis what more you need to add. Finally, make up the volume with nuclease free water as mentioned in your kit catalogue. And proceed to cDNA synthesis.

Dear student if you find the answer convincing please do give the feedback.