WEEK FOUR (February 6 - 10)

We have been given the DNA encoding all 11 genes and we need to get the DNA into cells, verify the inserts, freeze down glycerol stocks, enter them into GCAT-alog, and send some cells to our colleagues at MWSU.

Davidson Protocols

Everyone should register for a GCAT-alog account.

Friday February 3
transform JM109 plate on appropriate media. (Confirm with UW team)
take about 30 minutes

Saturday February 4
take out of incubator put in fridge
take about 15 minutes and must be done before 9 am

Sunday February 5
pick colonies from each plate and put into 2 mL appropriate media
take about 30 minutes

Monday
Do minipreps but SAVE THE CELLS on plates takes about 1 hour

Tuesday
set up digestions, run 30 min to 1 hr and put in fridge Determine insert sizes from Registry

Wednesday
determine the percent agarose needed for the inserts pour and run gel(s) then photograph gel(s) verify insert sizes (get this from Registry)

Thursday
grow overnight cultures of clones with correct inserts sizes pick from plate and put into 2 mL of appropriate media

Friday
take cells out of incubator freeze down cells and enter into GCAT-alog.