Receptor Experiment 1

To Do:

Friday:

• Nanodrop pENTR (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
• 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)
• PCR (possibly)

June 9

Plan of Action

 - Build hef1+lilrb2

  - design cloning strategy 

  - acquire promoter

  - acquire pdest 

  - build pentr lilrb2 

   - resuspend gblocks 

   - acquire gg domd

   - run a golden gate

Questions: 

1. How do we turn a gblock into an entry vector?
2. Can we get beta-amyloid in both monomer and oligomer form instead of just monomers that we would have to oligomerize?
3. Do we need the cells to be expressing GFP?

June 10 

Answers:

1. We will work on that later today
2. No preference
3. Makes for pretty pictures and not hard to do since we're already transfecting a plasmid

June 11

6/12/14

GoldenGate table: