ToDoTo Do:
Friday:
- Nanodrop pENTR
- Make 5 femptomolal working solution
- (was not useful)
- Transform pENTR into bacteria
- Incubate over night
- (5pm) Add Kan, incubate GGDonr liq culture (16hrs)
Saturday:
- Remove GGDonr liq culture from incubator (9am)
- Make cell stock, store at -80 degrees
- Put rest of cell culture in 4 degrees
Monday:
- Make empty liquid cell culture to transform DNA into (~16hours)
- Remove pENTR cultures from incubator (10:30am, Shinjini)
- Pick cells from pENTR colonies, put in liquid culture, put in incubator at 12pm
- Pick cells from pENTR colonies, put in liquid culture, put in incubator at 5:30pm (16 hours)
Sunday:
- Take pENTR colonies from incubator. Make cell stock. Put in fridge and freezer respectively. (10 am). Did not work.
...
Monday:
- Miniprep GGDonr (is that a good idea?)
- Pick Cells again and incubate (at 6pm)
...
Tuesday:
- Miniprep pENTRs
- Make 5 femptomolar working solution
- LR (~ 16 hours min, 24 max) Monday
- Miniprep GGDonrPCR (possibly)
June 9
Plan of Action
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| Making LilrB2/PirB expression vectors | Start Date | Progress/condition/next steps | Completed? (Date) | Result | Location of part (if needed, I don't know) | ||
|---|---|---|---|---|---|---|---|
Making pENTR from gBlocks: We have: We need: Golden-gating will result in: | 6/12 | GG mixtures in thermocycler GGDonr cell (culture?) in incubator Will make GGDonr bacterial stock today (6/13) | fridge (in case transformations did not work) | ||||
After that, Gateway (LR): Same for PirB We will get: 12_Hef1a_LilrB2 |
6/12/14
GoldenGate table:
...