Tag: ampicillin

Really quiet day in the lab today was…

Deborah / / Uncategorized

Really quiet day in the lab, today was the first day of my Uros, plasmid stability experiment. Started the day by growing E.coli C2110 to an OD of 0.35, tough to guess when this happens, 0.38 was the final reading. Whilst waiting for the cells to grow i then innoculated another flask of LB broth with C2110 and placed it in the 37C shaker.The next task was to make the cells competant and then transform them, using the plasmids pOG04 NJC and pOG04 DRW. The final step was to innoculate an LB agar plate (containing Ampicillin 50) and incubate overnight at 37C.

Hi all Interesting day today the broths i…

Deborah / / Uncategorized

Hi all,
Interesting day today, the broths i innoculated friday were ready today so i made them into stabs and slopes. Finally my workspace is clearing, made up a few more bijou jars ready for the end of the week when i can hopefully add Enterobacter spp and Citrobacter spp to my completed list. I then went through the cell competency procedure with Nicola and she assaulted my brain with maths (hate maths mondays), we then went over plasmid stability and what we were to achive this week. in preparation we made up 400ml LB agar, 400ml LB agar with chl10, 200ml LB agar with Ampicillin 100 and 200ml LB agar with Ampicillin 50. Tomorrow serial dilution!

Hey all I made some competent DH5 alpha…

Soph Marie Scanlon / / Uncategorized

Hey all!
I made some competent DH5 alpha E. coli cells today (which took 5 and a half hours to grow to the optical density of 0.350!) and used the colorimeter to see if they had reached the optical density . I have also transformed them so I can do alkaline lysis tomorrow! Still in the lab currently waiting as the transformed cells are currently in the incubator for 45 minutes then I need to spread plate them on to LB agar plates that have ampicillin in them and put them in a incubator over night. I’m using the same plasmids again (pALA1029, pOG4, pOG04, pOG4.003).
See you all tomorrow!
Soph 🙂

Busy day today Started the day going through…

Deborah / / Uncategorized

Busy day today,
Started the day going through wednesday and thursday’s results with Rachael, so she knows where we are with our cultures, and so we can plan what we’re doing next. Taught Hammad how to do water filtration, made up 950ml of M17agar base with 10g Lactose suppliment (suspended in 100ml distilled water), both were autolaved before mixing. After lunch Rachael poured the plates. Made up 2800ml (7 x 400ml) of LB agar and poured plates with five of these. The other two were left for Nicola to add ampicillin and then pour the plates.I also made up 250ml of 0.85% w/v NaCl, this also needed to be autoclaved. Made up two stabs and slopes with the PS aeruginosa broth that i incubated overnight. Counted colony cultures from Sharon’s Brayfod water project.