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

Gel 72810.jpg

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)