User:Susan Schultz/Notebook/Experimental Biological Chemistry/2011/11/02

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Objective

In Progress

Run PCR Product on Gel

Transform PCR Product into E. Coli

Run additional AuNP synthesis reaction with fresh gold stock

Description

AuNP Synthesis

Reaction 1- 1mL 15.5μM BSA + 1mL 2.5mM HAuCl4 removing from heat every 30 minutes for 10 minutes

Reaction 2- 1mL 15.5μM BSA + 1mL 2.5mM HAuCl4 heating continuously (some fluctuation from opening oven door)

Protein Gel

Remove wax from PCR Product

Add 1μL of DpnI and heat in heat block at 37°C for one hour to digest non-methylated DNA

Prepare 1.2% Agarose Gel: Add 0.25 g of 0.01 g/mL of agarose, 25 mL of 1x tris base, acetic acid, and TAE buffer in flask. Microwave for 40 seconds.

Pour gel and allow to solidify

Transfer 10μL of the PCR product to second PCR and add 2μL of 6X loading buffer

Load solution into gel, along with ladder


Transformation

Prepare agar plate: 0.875 g LB, 0.7 g agar, and 35 mL of water.

As mixture cools add 35 μL ampicillin.

Add 5μL of PCR product to 30μL NovaBlue Competent E.coli in sterile PCR tube

Data

  • Add data and results here...

Notes

This area is for any observations or conclusions that you would like to note.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.



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