IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/17

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png iGEM Project name 1 Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Biofilm Track

Eric F. + Melody

Biofilm (100816M + 100816E) Growth Protocol

  • Using Melody's 3mL cultures in plastic tubes of RN4220 + 8325-4
  • Inoculated flat bottomed 96 well plate with 1% glucose TSB RN42220 and 8325-4 in the below arrangement
  • Both plates were done exactly the same, at different times
Biofilms 100816M + 100816E Arrangement
Columns
Rows 1 2 3 4 5 6 7 8 9 10 11 12
A X X X X X X X X X X X X
B X X X X X X X X X X X X
C X X C R R R 8 8 8 C X X
D X X 8 8 8 C C R R R X X
E X X C R R R 8 8 8 C X X
F X X X X X X X X X X X X
G X X X X X X X X X X X X
H X X X X X X X X X X X X
  • I think Will check with the lab book tomorrow
  • Note: C = control, R = 1% glucose RN4220, 8 = 1% glucose 8325-4
  • 100816M (Melody's plate) went in at 1722
  • 100816E (Eric's plate) went in at 1830

dspB Track

O/N culture for miniprep

  • 3T1,3T2,4T1,4T2 in shaker @ 37C @1632

QS Track

  • Noticed agrCA from distribution came from wrong kit plate (it is on plate 2 instead of 1).
  • Resuspended agrCA in 15 μL dH2O and then transformed into DH5alpha cells using UBC iGEM protocols.
  • Digested and Ligated agrCA (natural Staph form) with B0014 (terminator) onto a PsB1C3 backbone. --Followed Biobrick protocols
  • Digestion time: 30 mins. Ligation Time: 50 mins.
  • agrCA was not purified using a spin column
  • Transform ligation mixture following UBC iGEM protocol. Use all 20 μL.