A Procedure for Freezing and Thawing Bacterial Cells - (Jan/21/2006 )
Hello. I'm a high school student who is doing a project which envolves me freezing and thawing bacterial cells. I need a procedure to do this, preferably using gylcerol. Can you help me in any way and please explain everything to me, as I'm only in high school.
for us to be able to help you, I think we will need more details about what you are trying to do.
could you kindly tell us as much as you can about your experimental design?
i.e., what type of cells? why are you freezing them? what are you hoping to show? why do you need glycerol?
there are many basic protocols for freezing cells. in order to determine which one is appropriate, we will need to know more.
If you want to demonstrate dthe importance of using glycerol to maintain theviability of your cells during the process.. you can compare thawing of cells with (10%) and without glycerol.. and count the colonies on LB plates considering the amount of cells that you froze.
Or demonstrate dthe best concentration of glycerol to use for the process
eg... using 100 ul gly+ 900 ul saturated cells in LB... 200 and 800.... 300 and 800.. etc etc
remember to countthe number of cell by making serial dilutions... e g. from the saturated before freezing take 1 ul and dilute in 99 ul of sterile water. and from this take anothe and dilute to another 99 ul... plate in lb plates the whole 100 ul, spreading uniformely using a spreader (maintain sterility)