Ambion
mMESSAGE mMACHINE T7 Transcription Kit
AM1344
I used this product to make mRNA for reporter assays without having to worry about any secondary effects of transcription. I choose this specific product after researching several different kits on the web. It's a kit to produce in vitro mRNA works well most of the time.
In vitro transcribe mRNA
Linear DNA
Linearize plasmid DNA. Add enzyme mix to linearized DNA. Put into thermocycler. I usually follow up with a polyA kit and then perform an RNA clean up
Make sure you have highly concentrated, clean DNA
I have hit or miss results with the output of the kit. I can put the same amount of linear DNA in, but get vary outputs of mRNA out.
N/A
When it works, it works well.
mRNA output can be variable
I tried several in vitro mRNA kits and this one generally works well, but you will sometimes have to repeat a reaction to get good output.