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Team:Brown/Notebook/July28
From 2010.igem.org
Wednesday, July 28, 2010
Ligations overnight
- Realized that the carrier plasmid of R0080 (pAraC) is still in the double digest product mix
- Need to separate so only the 149bp pAraC piece is put with the RBS_ChFP_TerTer plasmid
Plan: Run a gel (1%) of 18ul R0080 digest product mix + 4ul loading mixture next to 10ul ladder (1kb+)
- Gel showed a band ~150, cut and extracted. Nanodrop yield - 1.5(ng - could be ug, cannot read handwriting) per ul
pAraC digest product
M9 minimal media testing
- Saw growth in minimal media
- Made 1L of M9 minimal media (recipe on 7/26 page)
- Filter sterilized
L.Lactis overnight cultures for electroporation (from abandoned miraculin project)
50ul of saturated l.lactis in:
- 2.5ml MRS (2% glycine)
- 5mL MRS
- 7.5mL MRS
- 10mL MRS
to take O.D. in morning
Incubated @30C at 18:50.
pNoTat w/ WillRS ligation
- Transform 6ul of DNA into XL1s from Adrian
- Plated out on Amp+ LB
- Ran control transformations using control RFP
Overnight cultures for tomorrow
Made overnight cultures of:
- pLac/Mnt XL1B (amp+)
- CI (amp+)
- RBS34 (amp+)
4mL LB, 4ul amp stock
Tried to get competent cell stock (XL1B and BL-21)
