Hello all,
I have been using the mmessage mMachine SP6 and T7 kits from Ambion (ThermoFisher) to transcribe RNA. I get yields that are good enough for my work (~2-4ug/uL) and are in accordance with the manufacturer's description. The only issue is these kits are ridiculously expensive. They are ~$500/kit which gives 25 reactions, so, ~$20/reaction.
I have previously used a home-made kit long time ago (SP6, cap analog, dNTPs and RNase inhibitor) and have been able to get RNA although I had never quantitated it. I'm wondering if anyone has any experience with both methods and if there are large differences in yields and quality. I'm also wondering how cost effective the home-made version would be. TIA.
Ive used the kit based IVT products from Ambion/Thermo, they did work wonderfully. Cost wasn't an issue, however, so that is why we chose the kit versus home brew, in spite of the fact that it is relatively simple to do with off the shelf / catalog products.
The huge advantage (for kits) is the troubleshooting and support you can get if anything went wrong. You have one vendor to go to for troubleshooting. If your homebrew method gives you unexpected results, you have multiple products from multiple vendors to troubleshoot. So if time and efficiency is your concern go kit based, but if cost is a concern and you have more time (or interns :) ) you should certainly go homebrew.
Thank you Craig Silver. In my case, both cost and time are at a premium. Even though I understand that maybe in the long run the kits might be a better way to go in terms of reproducibility and tech support, I'm trying to see if I can get that kind of reproducibility with off the shelf products. Thanks for the input either way.
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