Correct kit for qPCR on Illumina preps?

Heisman · 06-07-2011, 10:35 AM

Greetings all,

We would like to start doing qPCR on our library preps, which are generally paired-end for the HiSeq2000. Is there a specific kit out there for this purpose? The following kit is stated as being for the GA: http://www.genomics.agilent.com/Coll...il&PageID=2053

If you guys use reagents not found in a kit, what do you use and how do you design your primers? Thanks a lot!

By the way, I searched through thread titles but the most promising threads were from September of last year, so I'm not sure they are still relevant.

kwaraska · 06-08-2011, 08:27 AM

There is a kit from Kapa for library quantification that works well.

We are using a homemade kit using Kapa SYBR reagents, primers, and PhiX as a standard curve. We tried the Kapa kit and it was just as good as ours, but ours is less expensive.

Heisman · 06-08-2011, 08:34 AM

Quote:

Originally Posted by kwaraska

There is a kit from Kapa for library quantification that works well.

We are using a homemade kit using Kapa SYBR reagents, primers, and PhiX as a standard curve. We tried the Kapa kit and it was just as good as ours, but ours is less expensive.

Thanks for the reply! How do you design your primers for the paired-end preps? I'm a rookie at qPCR so please excuse my ignorance.

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06-07-2011, 10:35 AM	#1
Heisman Senior Member Location: St. Louis Join Date: Dec 2010 Posts: 535	Correct kit for qPCR on Illumina preps? Greetings all, We would like to start doing qPCR on our library preps, which are generally paired-end for the HiSeq2000. Is there a specific kit out there for this purpose? The following kit is stated as being for the GA: http://www.genomics.agilent.com/Coll...il&PageID=2053 If you guys use reagents not found in a kit, what do you use and how do you design your primers? Thanks a lot! By the way, I searched through thread titles but the most promising threads were from September of last year, so I'm not sure they are still relevant.

06-08-2011, 08:27 AM	#2
kwaraska Senior Member Location: Boston,MA Join Date: Nov 2008 Posts: 122	There is a kit from Kapa for library quantification that works well. We are using a homemade kit using Kapa SYBR reagents, primers, and PhiX as a standard curve. We tried the Kapa kit and it was just as good as ours, but ours is less expensive.