Low Concentration Library

hanco · 04-14-2017, 12:31 AM

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 · 04-14-2017, 08:34 AM

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 · 04-16-2017, 04:53 PM

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

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
ddRAD library low concentration	dBlatt	Sample Prep / Library Generation	3	07-03-2017 11:30 AM
Library prep/low concentration	Normal	Sample Prep / Library Generation	1	03-19-2015 03:12 PM
Wang, 'Low Cost Library Construction...': Low Concentration	MAdkisson	RNA Sequencing	2	02-19-2014 02:33 PM
low library concentration	biochembug	Illumina/Solexa	0	08-02-2012 06:13 AM
Low concentration GA library	Melanie	Sample Prep / Library Generation	4	08-04-2010 01:34 PM

04-14-2017, 12:31 AM	#1
hanco Junior Member Location: Japan Join Date: Jun 2015 Posts: 8	Low Concentration Library 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!

04-14-2017, 08:34 AM	#2
pmiguel Senior Member Location: Purdue University, West Lafayette, Indiana Join Date: Aug 2008 Posts: 2,315	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

04-16-2017, 04:53 PM	#3
hanco Junior Member Location: Japan Join Date: Jun 2015 Posts: 8	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