Team:Warsaw/Calendar-Main/29 July 2008

From 2008.igem.org

Revision as of 14:14, 9 October 2008 by Marcinoll (Talk | contribs)

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
next notebook entry

 



1. Separate transformant colonies (transformation from previous day) inoculated to liquid LB with kanamycin.
2. Inoculation of pACYC177+OmpA_alpha and pACYC177+OmpA_omega - liquid LB with kanamycin.

Piotr:

Checking if OmpA_omega_A_alpha gives ampicillin resistance.

Inoculation OmpA_omega_A_alpha from various IPTG concentrations (0, 0.1, 0.25, 0.5, 0.75, 1) into the same IPTG concentrations, but with various ampicillin concentrations(25, 50, 75, 100) in relation: 1:50.

Measurement of bacterial culture growth (OD) in the evening:
ampicillin concentration (μg/mL):IPTG concentration (mmol/mL):
00,10,250,50,751
251,5581,4691,5871,491,5661,311
501,4251,4351,5241,0550,9200,935
751.090,9891.4470.9710.9510.992
1000.090,6851.3781.0780.9770.992

Inoculation of OmpA_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75 (repeat is needed!!!)


Checking OmpA_omega_A_alpha and OmpA_A_alpha expression.
  1. Spinnign
  2. Suspending
  3. Adding of lysis buffer
  4. Boiling
  5. Putting into poliacrylamide gel
  6. Transfer onto nitrocellulose
  7. Blocking
  8. Anti-A antibody binding
  9. Washing
  10. Anti-rabbit antibody binding
  11. Developing with BCIP and NBT
(Zdjęcie żelu)