Urgent SAC 1 U4 help :D!

[WhoTookMyUsername]

Hi guys, i've got the first part of my SAC 1 tomorrow..

it was the GTAC sac with electrophoresis and bacterial transformation using pGLO

i'm struggling to think of some errors for the SAC, anyone got some ideas :O ?
thanks!

[Russ]

It would help if you described your experimental method but the most obvious errors for those two procedures would be; incorrect separation of fragments, incorrect loading of the gel and no plasmid insertion

[WhoTookMyUsername]

by incorrect seperation of fragments do you mean the DNA fragments? how would you fix this ?


yeah sorry, um
i can't really remember the experimental procedure as we did it last term and don't have any info ATM

but

with electrophoresis

there was 2 DNA samples, along with loading dye, dna ladder etc. we measured base length of those (approx)

dna insertion, we spliced araC (what is the function of this? ) ampR and GFP gene and put them into plasmids. (after PCR i think , or maybe PCR was with electrophoresis)

then used heat shock for competence .

we put ethidium bromid on electrophoresis gel

[Russ]

araC regulates arabinose metabolism in E. coli (same as the lac operon but it's for a different sugar)

by incorrect seperation of fragments do you mean the DNA fragments? how would you fix this ?

Yeah, if you chose an inappropriate gel or pH etc. but upon reflection it's probably not relevant for you because whoever was running it would have done that for you.

[WhoTookMyUsername]

does the araC gene effectively allow an easy way of switching GFP on and off?

lol are there any other errors XD, as plasmid insertion did occur, and gel was loaded correctly (assuming phd student was competent)