T7 RNA polymerase

Product information
This product is recombinantly expressed in Escherichia coli .
Storage buffer: 20 mM Tris-HCl, 100 mM NaCl, 10 mM DTT, 0.1% Triton X-100, 1 mM EDTA, 50% Glycerol, pH7.9.
Reaction buffer: 400 mM Tris-HCl, 60 mM MgCl2, 10 mM DTT, 20 mM spermidine, pH 7.9

Instructions
1. Please refer to the following reaction system
0.4 μL 25mM NTP Mix, 0.2-1 ug DNA template, 2 μL reaction buffer, 0.5 μL RNase Inhibitor (optional, 1 U/μL), 1 μL 0.1 M DTT (optional), 2 μL T7 RNA polymerase, use RNase Make up the volume to 20 μL with free water.
2.Reaction conditions
Incubate at 37°C for 0.5-1h. For fragments smaller than 300 nt, it is recommended to incubate at 37°C for 2-16 h.

Notes
1. After the reaction, use 2 U DNase I (RNase-free) at 37°C for 15 minutes to remove the DNA template.
2. Adding 0.02 U thermostable inorganic pyrophosphatase to the 20 μL reaction system can significantly increase the transcription yield.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.