Tag: serial dilution

As i expected pALA1029 pOG04 DRW and pOG4003…

Deborah / / Uncategorized

As i expected, pALA1029, pOG04 DRW and pOG4003 have once again shown no growth, almost in sympathy the serial dilution i performed yesterday yielded poor results. The spread plates only showed colony growths of 1-14, as i was looking for colonies around 100, i decided that it was not worth replica plating. I have still carried out a serial dilution from the overnight broth cultures and spread plated them onto LB agar plates and left them to incubate at 37C overnight. I do not expect any growth based on todays results but one can hope!

Another frustrating day in the lab Grew pNC0412B…

Deborah / / Uncategorized

Another frustrating day in the lab!!
Grew pNC0412B to an OD of 1 then performed a serial dilution (A,B,C,D -1to-5.5), I innoculated a fresh 10ml LB broth with 100 microlitres from dilution -4 and placed into the 37C shaker overnight. I then spread plated 100 microlitres from dilution -5 onto a fresh LB agar plate, I also spread plated 100 microlitres from dilution -5.5 onto a fresh LB agar plate, then placed these into the 37C incubator overnight. I would have also done this for pOG04 NJC but it refused to grow, after 6 hours in the 37C shaker only an OD of 0.01 was achieved, i abandoned this set of broths. The plates i hoped to grow for today containing the plasmids pALA1029, pOG4003 and pOG04 DRW also showed no growth. I have tried again today so hopefully tomorrow will be a better day. I seriously think C2110 is the work of the devil!

Hi all counted yesterdays plates and replica plated…

Deborah / / Uncategorized

Hi all,
counted yesterdays plates and replica plated pOG4 onto LB and LB containing amp100, then i counted and replica plated pnc0412B onto LB and LB containing chl20, then i incubated them overnight at 37C. My next task was to complete the second serial dilution and spread plated 10-5.5 and 10-6 onto LB agar plates with no antibiotics. Next i checked on pOG04NJC and pnc0412B – NO GROWTH!!! I am sure they know its monday! Having Checked with Ross williams he thinks it could be the plasmid at fault. Tomorrow i try again.

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!

Hi Had a good couple of days have…

Deborah / / Uncategorized

Hi,
Had a good couple of days, have been having trouble with growth – not yesterday each plate had 500+ colonies, think i need glasses now!! Today was an easy day too, counted yesterdays T end plates and recorded the results, then counted the plates from yesterdays serial dilution T0 plates and replica plated the ones with around 100 colonies, once again there was good growth so i replica plated ones with around 200 colonies.

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).