Low Concentration Library

[hanco]

Hi,

I made libraries using Colon and Lung Cancer Research Panel and quantify them using qPCR. I was wondering why some of the libraries have less concentration than the others. I made libraries from serum DNA, the starting material is 50 ng and made all the libraries at the same time.
Can someone help me with the answer? I really appreciate it.

Thank you!

[pmiguel]

Hi Hanco,
There is no single answer to your question. Basically every step in your protocol has the potential to cause library construction to occur sub-optimally or even fail altogether. Also many methods of quantitating the amount of input material will be inaccurate. So the actual amount of material used for library construction may be highly variable.

--
Phillip

[hanco]

Hi Philllip,

I thought perhaps the quality of DNA that I used to make the libraries already damaged from the beginning or my technical error.
Anyway, thank you so much for your answer.

Hanco