Lysis buffer preparation

Use this category for general molecular biology questions that don't fit specifically into any of the categories above, including software questions

Moderators: r.rosati, mdfenko

Lysis buffer preparation

Postby contaminantRNA » Aug 02 2017 7:10 am

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 required lysis buffer can be 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 lysis buffer. The RNase inhibitor I will order is 100 µL at 20 U/µL.

Please walk me through how to prepare the lysis buffer. Thanks.
contaminantRNA
newcomer
newcomer
 
Posts: 1
Joined: Aug 02 2017 7:00 am

Re: Lysis buffer preparation

Postby mdfenko » Aug 03 2017 6:27 am

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).
talent does what it can
genius does what it must
i do what i get paid to do
mdfenko
ModSquad
ModSquad
 
Posts: 1456
Joined: Jan 24 2006 2:39 pm
Location: Staten Island, NY USA


Return to General Methods

Who is online

Users browsing this forum: No registered users and 1 guest