IGEM:MIT/2007/Notebook/2007-7-24: Difference between revisions
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(New page: ==AGENDA== *Plate the Mer/I13500 3A transformation after overnight ligation (and grow) *Check if sequence results are available from Monday (length was correct) *Second polystyrene assay (...) |
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**Harass the people AK e-mailed OR design and order? | **Harass the people AK e-mailed OR design and order? | ||
*If T7 antibody comes in...test! | *If T7 antibody comes in...test! | ||
===Transformation of CPX (A+,Cm+), FhuA (K+) and pUC18 (A+) into Barry's BL21 cells (more protein!)=== | |||
*Obtained pUC18 and BL21 courtesy of Barry | |||
*Cell volume: 50 µl | |||
*DNA mass: 50 to 0 ng | |||
*For spreading on plate: 300 µl and let dry | |||
*NOTE: to help plates absorb liquid bacteria, leave plates out overnight after making them before putting them in fridge (dries them out) | |||
*"BL21 trans. w/ pUC18 (A+)": pulled out 55 µl of pUC (at 1 ng/µl of stock) and added to cells | |||
*"BL21 trans. w/ FhuA (K+)": diluted 1 µl of stock into 2 µl total; pulled out 1.5 µl and added to Bl21 | |||
*"BL21 trans. w CPX (Cm+ A+)": diluted 1 µl of stock into 4 µl total; pulled out 1.5 µl and added to BL21 |
Revision as of 13:37, 24 July 2007
AGENDA
- Plate the Mer/I13500 3A transformation after overnight ligation (and grow)
- Check if sequence results are available from Monday (length was correct)
- Second polystyrene assay (JH and TTP?)
- CPX, PBS with different concentrations and controls
- Suggested: one person per plate, pipet to the edge, more washes, harder aspirations
- Sequence the FhuA (pHIE) with CORRECT primers (which already exist?)
- Prepped plasmid exists (AL, 7/23)
- EC_20 requests
- Harass the people AK e-mailed OR design and order?
- If T7 antibody comes in...test!
Transformation of CPX (A+,Cm+), FhuA (K+) and pUC18 (A+) into Barry's BL21 cells (more protein!)
- Obtained pUC18 and BL21 courtesy of Barry
- Cell volume: 50 µl
- DNA mass: 50 to 0 ng
- For spreading on plate: 300 µl and let dry
- NOTE: to help plates absorb liquid bacteria, leave plates out overnight after making them before putting them in fridge (dries them out)
- "BL21 trans. w/ pUC18 (A+)": pulled out 55 µl of pUC (at 1 ng/µl of stock) and added to cells
- "BL21 trans. w/ FhuA (K+)": diluted 1 µl of stock into 2 µl total; pulled out 1.5 µl and added to Bl21
- "BL21 trans. w CPX (Cm+ A+)": diluted 1 µl of stock into 4 µl total; pulled out 1.5 µl and added to BL21