Lysis buffer preparation

Go to the profile of contaminantrna
Aug 02, 2017
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Hey guys,

I've been away from molecular biology for a while and I just want some help with calculations. I need to prepare a lysis buffer for single cells. The guidelines I have are as follows: The requiredlysisbuffer canbe made by combining 0.2% (vol/vol) Triton X-100 and 2 U/µl RNase inhibitor.

I need to freeze my cells in 2.3µl lysisbuffer. The RNase inhibitor I will order is 100 µL at 20 U/µL.

Please walk me through how to prepare the lysis buffer. Thanks.

1 Comments

Go to the profile of mdfenko
mdfenko 5 months ago

the secret to preparing solutions from stocks is:

c1v1=c2v2

most of us don't have the ability to measure out the volumes required to prepare 2.3ul of the solution you require. we prepare larger volumes (0.5-1ml).

calculate how much of each component you require to get the concentrations you state, add together and adjust to the final volume.

(i don't normally do the math for posters but i'm feeling generous today) for 0.5ml final volume, you need 1ul triton (1/500th=0.2%) and 50ul inhibitor (the stock is 10x your final concentration).