User:Omar Choudary/Notebook/Omar Choudary CHEM-571 AU-2011/2012/2011/11/02

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Objective

  • Perform gold nanoparticle(AuNP) synthesis using 15.5 μM Bovine Serum Albumin (BSA), 2.9 mM Chloroauric Acid (HAuCl4), and H2O.
  • Transform mutated DNA synthesized from Plasmid PCR reaction performed on 11/1.

Description

  • AuNP Synthesis:

1. Mixture created in the following order: 1 mL BSA (15.5 μM), 1 mL HAuCl4 (2.9mM), 8 mL H2O.

2. Placed in oven, removed every 30 minutes for 10 minutes over the course of 2 hours.


  • DNA Transformation:

1. LB/Agar Plate

A. Mixed 0.875g LB, 0.7g Agar, 30 mL H2O

B. Autoclaved for 90 min

C. Added 35 μL Ampicillin (Antibiotic), 35 mL H2O

D. Poured into petri dish until bottom of dish is fully covered

E. Solution left to dry until solid in petri dish


2. Prepare the cells:

A. Placed DNA solution from PCR on ice

B. Added 5 μL DNA solution to 30 μL of E.Coli cells

C. Incubated cells on ice for 30 min

D. Moved cells to heat block set at 42 degrees for 30 seconds (Heat Shock)

E. Cells then returned to ice for 5 min

F. 250 μL SOC media added to cells

G. Incubated in shaker set at 37 degrees celsius for one hour

Data

  • Molar ratio of Au/BSA - 187.
  • Solution was clear with purple protein fibers forming.
  • Compared to the experiment performed on 11/1, the fibers were spread throughout the entire test tube as oppose to forming a clumpy aggregate.

Notes