Hi everyone
I am planning to use the Qiagen RT2 Profiler PCR array for signaling pathways. Is it really necessary to use the RT2 SYBR Green Mastermixes and the RT2 First Strand Kit with the profiler array? Can I use Applied Biosystems SYBR Green PCR mastermix instead (Cat no. 4309155) as well as my already synthesized cDNAs? The platform is Roche Light Cycler 480.
Looking forward for some help. Many thanks.
We have successfully used non-Qiagen PCR master mixes with their PCR arrays. I suggest setting up a PCR plate that would have a few reactions: one or two Qiagen PCR assays (read primers; these are the same ones used in their arrays; if you have them separate in a tube) mixed with a recommended Qiagen SYBR Green vs. third-party SYBR Green. This is going to be your ultimate validation. But you lose any warranty this way. Good luck.
I have seen this question asked a ton of times.
Use the Qiagen/SuperarrayBiosciences RT and mastermix because it has built in controls combined with the profiler plate to tell if at what point you have a failure qPCR these are in wells H6-H12.
http://www.sabiosciences.com/pcrarraycontrols.php
Hi Kenneth, the use of Qiagen qPCR mix has nothing to do with all the controls. HKGs, gDNA contamination and negative controls can be detected with any master mix. To make RT efficiency control work all you need is to use a Qiagen RT kit. Once you have cDNA for the synthetic RT control RNA in your sample made by the Qiagen RT kit, you can use any qPCR master mix to detect it. The primers are already on the plate. If you want to avoid using Qiagen RT kit, you may choose much cheaper RT efficiency kits and uncouple this control from your qPCR plate. The only thing you are losing by not using Qiagen kits in all steps is their warranty.
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