User:Allison K. Alix/Notebook/Thesis Research/2013/07/12
DNA/AuNP attachment | Main project page Previous entry Next entry |
To Do
Calculating DNA and AuNP concentrations
20mM(500uL) = 268mM(x) x=37.3uL sodium citrate to be added to AuNP/DNA solution (500-37.3)/2 = 231.3uL each of DNA and AuNP to be added.
x = 2.226uM <-----this is the concentration prior to mixing with AuNP/sodium citrate 2.226uM(500uL) = x (250uL) x = 4.45uM <---this is the concentration prior to mixing with 250uL 100mM DTT in 4% TEA Concentration of stock DNA = 53.1uM 53.1uM(x) = 4.45uM(250uL) x = 21uL in 229uL 1X PBS Preparation of 100mM Sodium Citrate Buffer
to prepare 100mM sodium citrate-HCl buffer: 294.1g/mol x 0.1 mol/L x 0.01L = 0.2941g in 10mL H2O adjust pH and volume accordingly Notes/Observations-AuNP's still seem to be aggregating even after using freshly prepared citrate buffer. We attribute this to a few things 1) Waiting 5 minutes for the AuNP and DNA to react before adding the sodium citrate 2) The concentration of sodium citrate in solution should actually be 10mM NOT 20mM (oops...) 3) The sodium citrate buffer prepared initially only contained sodium citrate and NOT citric acid. Fresh buffer contained both. To do on Monday, 7/15 1) Add AuNP/DNA/sodium citrate all at once and vortex immediately 2) Prepare 2 samples, one with 100mM sodium citrate buffer prepared today containing only sodium citrate and HCl and one with 268mM sodium citrate buffer prepared yesterday containing sodium citrate, citric acid, and HCl
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