Tag: transformation

Today may be Friday but because of the…

Deborah / / Uncategorized

Today may be Friday but because of the bank holiday monday, i am treating today like a monday!!
I am growing two batches of C2110 up to an OD of 0.35. Then i will perform cell competancy and transformation. Normally i would put the spread plates in the 37C incubator overnight but because of the bank holiday i am placing the plates in the 25C until tuesday. Hopefully there will be some growth. If all goes well Tuesday i will be working with pNC0412B, pOG04NJC, pALA1029 and pOG4003.

Today has been an almost perfect day in…

Deborah / / Uncategorized

Today has been an almost perfect day in the lab.
pOG4 and pNC0412B have grown quite nicely to an OD of 1 (0.8-1), I have performed a serial dilution on both and left the innoculated broths overnight in the shaker at 37C, the spread plates of 5 & 5.5 are in the 37C incubator. Also today i innoculated another flask of LB broth with C2110 and grew to an OD of 0.35. Then i carried out cell competancy then transformed them, using the plasmids pOG04 NJC and pNC0412B. I spread plated pOG04NJC onto an LB agar plate containing amp50 and spread plated pNC0412B onto an LB agar plated containing chl10. these were then placed in the 37C incubator overnight. Busy day tomorrow as agars, broths, velvets and NaCl need to be made-up/autoclaved, as well as tomorrows tasks. Early start i think!

Groundhog day Today is still Monday no overnight…

Deborah / / Uncategorized

Groundhog day!!!!
Today is still Monday, no overnight growth again. Think C2110 is mocking me. I have repeated everything once again but i have tried to trouble shoot as to why nothing is working. Once i made thre cells competant i plated a sample onto LB agar plate and a sample onto LB agar with amp50. Then i continued on to the transformation stage, i then plated a sample onto LB agar plate and onto LB amp50 agar plate. I have also restreaked a fresh nutrient plate with C2110. Hopefully things will grow overnight, if not maybe i can work out at what stage things are going wrong.

Hey Yesterday i grew C2110 E coli to…

Deborah / / Uncategorized

Hey,
Yesterday i grew C2110 E. coli to an OD of 0.35. Three hours later!! I made the cells competant, then i transformed them, leaving them in the 37C shaker for 45 minutes. Once this was complete i spread plated 0.1ml onto an LB agar plate containing amp50 and left the plates to incubate overnight at 37C. Unfortunately this morning NOTHING had grown. very frustrating, so today is now monday and i’m repeating everything again!!