Hello , I'm planning to perform an RNase A treatment before IP any suggestion for the incubation time at 37C or how much I should add RNase?

20 April 2021 1 8K Report

I'm planning to use RNase A/T1 Mix ( 2 mg/mL of RNase A and 5000 U/mL of RNase T1)

Hi Razan,

Looks like you're using something similar to ThermoFisher's A/T1 mix:

https://www.thermofisher.com/document-connect/document-connect.html?url=https%3A%2F%2Fassets.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2FMAN0012005_RNase_AT1_UG.pdf&title=VXNlciBHdWlkZTogUk5hc2UgQS9UMSBNaXg=

There you can see they suggest to use 20 µL mix per 1 mL.

Incubation time is a bit unclear, but also depends on the input.

Considering you want to perform IP and have low'ish sample volumes I'd feel 15 minutes would be sufficient. 30 minutes the most.

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