QuantiTect Primers with Roche SYBR - ....can you mix and match? (Feb/18/2009 )
I need to run some gene expression assays fairly quickly and the QuantiTect Primer Assays available from Qiagen appear to be an easy way to get data fast. I can buy primers for my genes of interest and an internal control housekeeping gene without having to optimise We have a LC480 in the lab and a good stock of Roche SYBR green master mix, so I'm a bit reluctant to buy the equivalent from Qiagen for just a few experiments (in the long term I'd like to optimise and run everything using our Roche kits, etc).
My question is, has anyone ever tried using Qiagen primers and SYBR green master mix from another company? Qiagen say I won't have accurate expression if I do this, but will it really make that much of a difference?
Thanks in advance,
I wish I knew the answer to this question,...unlike you, I'm going to buy the rotorgene RT-PCR kit from Qiagen, but order the primers from another company. I didn't know we could also buy primers from Qiagen. I asked the supplier, but he said Qiagen doesn't have any primer design or synthesis service. I'm in Singapore, so they might not really know. however I read the Gene Globe search engine on Qiagen website, seems that you need to buy those primers sets in combination with your PCR kits made for your PCR machine format (rotor-based or block-based)....
just now I searched for my gene of interest to give me primer sets, but it doesn't say what is inside the tube or doesn't say anything about the primer sequences.
I honestly don't understand why you buy Qiagen material for LC480. because Roche has its own kits ( I read you will optimize for Roche in the future, but still don't understand why). I think Roche has its own primer design and stuff too.
Yup, I regularly mix and match my primers and SYBR green - never had a problem. When I first started real-time I was worried I shouldn't do this, however I've tested a few SYBR mixes now and I found the Roche mix worked best with my samples. Personally, I think the companies just say this so you'll use all their reagents and spend more money with them.
Hope that helps!