I'm doing the mutagenesis process, but i dont get it

[mutagenesis]

I'm working on mutagenesis with deletions in the OS9 plasmid.

So I run a pcr to cut its specific area, do a transformation, wait overnight for cells to grow, pick colonies, rotate overnight in agar, miniprep or maxiprep, and then run the dna gel electrophoresis.

but what am I actually doing? I don't really understand what im doing. Can someone explain each step and why I do it?

[jiajia1987]

I suppose you are performing a deletion PCR (you need to confirm this with me)? If this is the case, you can refer to:

PCR-mediated deletion of plasmid DNA
by: M. Hansson, K. Rzeznicka, M. Rosenback, M. Hansson, N. Sirijovski
Analytical Biochemistry, Vol. 375, No. 2. (15 April 2008), pp. 373-375.

After the PCR, transformation is done so that the plasmids (with the deleted portion) can be replicated inside the bacteria. The bacteria is grown on agar plates and colonies (from bacteria that has been successfully transformed and which contains plasmids with the desired antibiotic resistance) are picked to grow in broth. This broth is incubated overnight, following which a miniprep or a maxiprep is carried out to isolate the plasmids with the deleted portion.

I only told you very briefly for each step. If you need more help, just ask more specific questions for us to answer. =)