Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression/24 July 2015"

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AGENDA FOR JESSIKA:

1. Finish yesterday's purification and save the fractions in 4 degrees. If possible, repeat the purification using another sample of raw cell ly sate. Don't forget to use the elution in column steps, and recharge and re-equilibrate the buffer before finishing!

2. Create a 1L culture of the 9-mer from one of the starter cultures. MAke sure you OD the starter culture and record the OD in a notebook! You can prepare the LB in a 2L flask from the side of the sink and autoclave it in the same flask. Once cool, add the chloramphenical (1mL) and then one of the starter cultures.. Make you add the IPTG after 3-4 of inoculation (check the OD for 0.6-0.6, and let it run over night).

3. On saturday, spin down the cell pellets from the overnighted culture.

OD Checks

After 17 hour start-up culture inoculation: 0.376

4 hr incubation, before IPTG added: 0.55

NOTE: Only around 4.2mL of IPTG stock solution left, so added 0.8mL less than originally planned.

1st hour after IPTG addition: 0.65

2nd hour after IPTG addition: 0.82

3rd hour after IPTG addition: 1.10

4th hour after IPTG addition: