Fast way to get plasmid for PCR - (Jan/11/2008 )
Hi everyone,
I am picking clones from the plate to screen which one has the right inserted plasmid by grow them in LB and then do PCR.
I heard that with only boiling the bacteria but not miniprep we can get some plasmid DNA which is not pure not concentrated but is good enough for PCR. I never do that and do not know the detail. Does anyone do that? If so, how is it?
Thank you.
Dan
it's quite easy. Take plate with your clones and a new labeld plate.
With a fine tip, just pick up one clone, tap the tip on teh second plate, and put the tip in a pcr tube containing 10µl of ddH20. let it for 1to 2' to let bacterias detach from the tip.
repeat the procedure with as many clones as the number you want to test.
Wen finished, swirl gently the tips immerged in the ddH20 and put 15µl fo your pcr mix (including the taq).
Go for your pcr cycle with a first 5' 95° boiling/denaturing step
.
With a fine tip, just pick up one clone, tap the tip on teh second plate, and put the tip in a pcr tube containing 10µl of ddH20. let it for 1to 2' to let bacterias detach from the tip.
repeat the procedure with as many clones as the number you want to test.
Wen finished, swirl gently the tips immerged in the ddH20 and put 15µl fo your pcr mix (including the taq).
Go for your pcr cycle with a first 5' 95° boiling/denaturing step
.
Thank you fred. I am going to try that now!