User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2011/09/27: Difference between revisions

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Chloroauric acid --> 17.0 mg in 10 mL of water --> 4.3 mM
Chloroauric acid --> 17.0 mg in 10 mL of water --> 4.3 mM
BSA --> 10.3 mg in 10 mL of water --> 15.4 uM
BSA --> 10.3 mg in 10 mL of water --> 15.4 uM


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Also today, we will be transforming our DNA from last week into cells. First we have to digest the wild-type dna with an enzyme called DpnI (I will explain this in class). Then we have to follow a procedure for plating our cells.  
Also today, we will be transforming our DNA from last week into cells. First we have to digest the wild-type dna with an enzyme called DpnI (I will explain this in class). Then we have to follow a procedure for plating our cells.  


1) Make LB/agar plates with antibiotic (will describe in class)
# Make LB/agar plates with antibiotic (will describe in class)
2) Place sterile tube and your DNA (post DpnI reaction) in ice bucket for 15 minutes while thawing and cells.
# Place sterile tube and your DNA (post DpnI reaction) in ice bucket for 15 minutes while thawing and cells.
3) Add 5μL of your DNA and 40μL of cells to the bottom of the tube.
# Add 5μL of your DNA and 40μL of cells to the bottom of the tube.
4) Incubate on ice for 30 minutes
# Incubate on ice for 30 minutes
5) Add 250μL of SOC media (I will supply this)
# Heat shock your DNA/Cells at 42C for 30seconds
6) Incubate in shaker at 37°C for 1 hour
# Incubate on ice for 5 minutes
7) Spread 100μL of cells on your LB/agar plate
# Add 250μL of SOC media (I will supply this)
8) Store plate (inverted) in 37°C oven overnight.
# Incubate in shaker at 37°C for 1 hour
# Spread 100μL of cells on your LB/agar plate
# Store plate (inverted) in 37°C oven overnight.


==Description==
==Description==

Revision as of 11:14, 27 September 2011

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Objective

Starting concentrations for Chloroauric acid and BSA solutions:

Chloroauric acid --> 17.0 mg in 10 mL of water --> 4.3 mM

BSA --> 10.3 mg in 10 mL of water --> 15.4 uM

I want you to use these stocks to make solutions for synthesizing nanoparticles. Use the same final concentrations that you used on the 7th. Also, use the same buffers that your groups were assigned to. Tomorrow we will all use water for the synthesis. I will give you directions for how we are conducting this experiment at the beginning of class.


Also today, we will be transforming our DNA from last week into cells. First we have to digest the wild-type dna with an enzyme called DpnI (I will explain this in class). Then we have to follow a procedure for plating our cells.

  1. Make LB/agar plates with antibiotic (will describe in class)
  2. Place sterile tube and your DNA (post DpnI reaction) in ice bucket for 15 minutes while thawing and cells.
  3. Add 5μL of your DNA and 40μL of cells to the bottom of the tube.
  4. Incubate on ice for 30 minutes
  5. Heat shock your DNA/Cells at 42C for 30seconds
  6. Incubate on ice for 5 minutes
  7. Add 250μL of SOC media (I will supply this)
  8. Incubate in shaker at 37°C for 1 hour
  9. Spread 100μL of cells on your LB/agar plate
  10. Store plate (inverted) in 37°C oven overnight.

Description

  1. Add experimental record here. Include what, how, and why...

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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