I am using the MEGAscript RNAi Kit for synthesizing dsRNA for RNAi. I ran my reaction product on a denaturing gel and got a smear. Can you offer some advice?
If the RNA appears degraded (e.g., smeared), remove residual RNase from the DNA template preparation before in vitro transcription. Do this by digesting the DNA prep with proteinase K (100-200 µg/mL) in the presence of 0.5% SDS for 30 min at 50°C, and follow this with phenol/chloroform extraction. The RNase Inhibitor that is present in the transcription reaction can only inactivate trace RNase contamination. Large amounts of RNase contamination will compromise the size and amount of transcription products.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.
What is the recommended shelf life of the MEGAscript RNAi Kit (Cat. No. AM1626)?
The MEGAscript RNAi Kit (Cat. No. AM1626) has no specific expiration date. However, for our products without specific expiration date, we guarantee best performance for one year effective from the date of receipt, when stored as recommended.
Find additional tips, troubleshooting help, and resources within our RNAi Support Center.