dilution series

Hey all,
Yesterday i took the overnight broths for the two plasmids and completed another dilution series and spread plated the ones from -5.5 and -6. I incubated these at 37C overnight. Then i took out the T0 plates and counted the colonies, the ones that were around 100 i replica plated them onto an LB amp100 agar plate and then olnto an LB agar plate. These were then incubated overnight at 37C.
Today started out with me making more LB agar and LBamp100 agar plates, and making up 3x50ml LB broth, ready for monday morning. Then i autoclaved my velvets ready for next week. Once this was completed the science could start. I counted Tend plates and replica plated the ones with around 100 colonies, same as yesterday i replica plated 1x LBamp100 and 1xLB agar. The last job of the day was to count the colonies from yesterdays replica plating and record them. I also re streaked my C2110 so that i will be working with a fresh culture on monday.

YAY!! got growth, finally can start. I took four colonies from each of my culture plates and made them up into broths using LB broth and adding ampiillin 100. Then i placed them in the shaker at 37C, until they grew to an OD of 1(0.8-1). then i set up a dilution series, labelling eppendorfs A,BC,D -1 to -5 adding 0.9ml NaCl and 0.684ml into the eppendorf marked -5.5. Once the broths reached an optical density of 1, 0.1ml was added to each eppendorfs with the exception of the -5.5 as 0.316ml broth was added. From the 10 -4 eppendorf, 100microlitres was innoculated into 10ml of LB broth and incubated overnight at 37C. The -5 and -5.5 dilution series were spread plated onto LB agar plates and incubated overnight at 37C. This was repeated for pOG04 NJC and pOG04 DRW plasmids. Cant wait for tomorrow.

Tag: dilution series

Hey all Yesterday i took the overnight broths…

YAY got growth finally can start I took…