Thursday, September 22

From 2011.igem.org

Steve, Dr Liz present

We picked single colonies from the pET-Taq (4A4) in the 10-alpha strain of E. Coli growing on one of the two plates Dr Burkett brought for us. Made 5 samples for incubation in the 37° shaker:

Each of the first 4 tubes contained 3ml of the liquid growth medium, while the fifth tube only had 1.5ml. We lightly touched 4 separate colonies, putting those into the first 4 tubes. We dropped the tip used to pluck each cell sample into its respective tube. For the 5th tube we again touched the same colony that we touched for Tube 4. We are curious to see if there is a difference in growth between samples 4 & 5. Our media was LB enhanced with 50mM d-glucose (also called glucose or dextrose)..

We refreshed our ROOM-Temp TSA plus AMP plates (given to us by Cheryl Brown) with more AMP. We used 100ul on each plate, spreading with an ethanol-dipped and flamed glass wand. They absorbed the liquid sitting gel-side-down for just under an hour credit we taped them back into their plastic sleeve, gel on too, and put in our box in the cold room.

We set up 4 positive control PCR rxs using most of the remaining sample of purified pET-Taq (4A4) as template. We created a tube of 0.06 ug/ul template by mixing the full-strength pET-Taq with the 4A4 that had been diluted to a 1/100 concentration.