Tag: e.coli

Hi all Catching up with last week the…

Deborah / / Uncategorized

Hi all,
Catching up with last week, the plates i innoculated to try and trouble shoot the non growth issues i have been experiencing, showed that there is growth at each stage so at least i can rule out human error, yay! The strain of E.coli i am using (C2110) may be the issue as it is taking 6-7 hours to grow to an OD of 0.35, i would expect the growth to be in the region of 3 hours+. Leaving the plate with no growth on my desk to show mike the next day, miraculously i had fantastic growth overnight. I have already generated data for the plasmids pALA1029, pOG04 NJC, pOG04 DRW, pOG4003 and i am now starting with pOG4 and the Chloramphenicol resistant pNC0412B, fingers crossed all goes well.

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 Started day by completing my serial…

Deborah / / Uncategorized

Hi all,
Started day by completing my serial dilutions from yesterdays broth culture, I spread plated the dilutions of 10-6 and 10-6.5 on LB agar and incubated them overnight at 37C. Then taking the eight plates from yesterday I counted the colonies and chose the four with around 100 colonies. I then replica plated each of these onto a further two LB agar plates (one containing Ampicillin 100). These were then incubated at 37C overnight. I then also broth cultured another confirmed strain of E.coli (e0791).

Hi Started the day Gram staining E coli…

Deborah / / Uncategorized

Hi,
Started the day Gram staining E.coli a strain i dont have, this is another cultue confirmed so i then made a broth which i am incubating overnight at 37C. Spent most of the day waiting for cells to grow to an OD of 1, though 0.8 – 1.5 would be within range. Nicola went through serial dilution with me and showed me how to do the first set, i then completed the following three, broth culturing -4 dilution and spread plating -5 and -5.5 dilutions onto LB agar plates and incubated them overnight at 37C. Hopefully tomorrow we will have enough growth to count colonies (100 would be nice) then we can do replica plating. Had another Mike Shaw masterclass and debate (loving lab life xx).

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 🙂