Only mRNA containing 7-methyl guanosine (M7G) cap structure at the 5'end and a Poly(A) tail at the 3'end can be effectively expressed in eukaryotic cells. Capped RNA synthesis kit enables the production of a large number of RNA containing a 5' Cap structure in vitro, which is capped to the 5' end of RNA by the use of anti-reverse Cap Analog (ARCA).
ARCA structure:
Using the control template supplied with the kit, each reaction will yield approximately 60-70 μg of RNA.
Materials provided with the kit
The kit contains sufficient reagents for 10 reaction of 20 μL each. Each standard reaction yield up to 70 μg of capped RNA, 2300nt length, from 1 μg control template.
Component
10 rxn kits
Storage
5× Reaction Buffer*
40 μL
-20°C
2× ARCA/NTP mix
100 μL
-20°C
Enzyme Mix
10 μL
-20°C
Control Template(0.5 μg/μL)
10 μL
-20°C
DNase I (RNase-free, 1 U/μL)
10 μL
-20°C
Nuclease-free Water
1000 μL
-20°C
Ammonium Acetate Stop Solution
500 μL
-20°C
Lithium Chloride Precipitation Solution
500 μL
-20°C
Gel Loading Buffer
100 μL
-20°C
* Salts, buffer, dithiothreitol, and other ingredients; 5× Reaction Buffer store at -70°C may result in the formation of a white precipitate. If this happens, heat the tube to 37°C for 5 minutes and mix thoroughly to resuspend the precipitate.