Prepping and transformation - (Apr/24/2006 )
So same as my previous query, one more stupid query.
When you insert the ligated product into the bacteria and you want to have lot of this ligated product, you grow those bacteria overnight and open up to have the ligated product. I was wondering why this process is called as Prep?? Is this short-form of something?
Another thing which I guess I know but not too sure with my chemistry background.
I have been given a plasmid which my boss wants me to get into some bacterial strain, thats exactly as transformation, right????
All I planned was, to get that plasmid, add the bacterial strain that he wants me to put it in, put it on ice for 30 mins, heat shock at 42sec for 45 mins and then put LB medium to grow these cells for couple of hours and then plate this bacterial sample of LB plate - Quick word transformation. Am I right here??
I thought I am, so why did she not use only transformation and asked me to 'get this plasmid into bacteria blah blah' Does this phrase mean something different? She is not very approacheable
yeah, the term is transformation...perhaps she thought it would confuse you and that you didn't know what that meant?
you have to make "competent cells" before mixing them with your plasmid DNA in order for the transformation to be successful; I can tell you if you search this site you'll find many good protocols
also, when you 'prep' your plasmid, you are making a large-scale preparation of the DNA.
Thanks for your reply.
This is not the first time that this thing has happened. Its the everyday story to fool me around and make me scratch my head for no reason and then scream for working slowly. I don't know what kind of fun this is for her??