User:Floriane Briere/Notebook/CHEM-496/2011/11/02

From OpenWetWare

Jump to: navigation, search
Image:BDLlogo notext ir.png Project name Main project page
Previous entry      Next entry



Objective

Today we are going to perform two different experiments:

  1. We are going to perform some Gold NPs again using exactly the same protocol as yesterday (1st of November) but we are going to use a fresh gold solution.
  2. We are going to transform the mutate DNA we synthesized yesterday (1st of November) thanks to the PCR technique.

Description

  • Gold NPs synthesis:
  1. In this specific order, add 1ml of BSA stock solution (15.5µM) + 1ml of HAuCl4 stock solution (2.9mM) + 8ml of water
  2. Place in the oven for 2 hours; remove every 30 minutes and leave the solution at room temperature for 10minutes
  • DNA transformation
  1. Prepare the LB/agar plate:
    1. Add 0.875g LB + 0.7g Agar + 35ml H2O
    2. Leave the solution in the autoplate for 1hours and a half
    3. Add 35µL of Antibiotics (Ampicillin) + 35mL of water
    4. Put some solution into a plastic Petri dish
    5. Let the solution become solid
  2. Prepare the cells:
    1. Place the DNA solution in the ice during
    2. Add 5µL of DNA solution to 30µL of cells solution
    3. Incubate on ice during 30 minutes
    4. Place the solution at 42°C for 30 seconds to make a heat shock
    5. Incubate in ice for 5 minutes
    6. Add 250μL of SOC media
    7. Incubate in shaker at 37°C for 1 hour

Results

At the end of the Gold NPs synthesis, we obtained some purple fibers; they began to form after 1 hours in the oven and became darker after 1 hours and a half; the fibers are spread through the solution and don't form an aggreggate like yesterday.




Personal tools