User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/09/16

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Objective

To create more AUNP with lysozyme fibers as most of those from 9/15/15 failed to make fibers. Only one 5mL and four 1mL samples generated fibers. The other goal was to make 500 mL of 10 mM Tris buffer, pH 9 and 500 mL of 10 mM phosphate buffer, pH 7.

Description

The instructions for the day can be found here.

  1. Making the AUNP with lysozyme fibers
    1. We followed the same guidelines as we used for 9/15
    2. There were only enough reagents to make 4 1mL samples
  2. Making the buffers
    1. Tris buffer: 500 mL, pH 9, 10 mM
      1. Used buffer calculator to find the mass of Tris buffer required
        1. Selected Tris for molecular weight, entered 500mL for volume, and 0.010 for molarity. Calculator advised adding 0.6157 g of Tris.
    2. Phosphate buffer: 500 mL, pH 7, 10mM
      1. Used phosphate calculator to find amount of dibasic Na2HPO4 and monobasic NaH2PO4 required
        1. Entered 7 for pH, 10 mM for buffer strength, ___ for MW_Na2HPO4, and __ for MW_NaH2PO4. Calculator advised adding 0.39816g of monobasic sodium phosphate and 0.36831g dibasic sodium phosphate

Results

  1. AUNP with lysozyme fibers
    1. No fibers were created, all solutions turned purple but no fiber formation
  2. Buffers
    1. Tris buffer: 500 mL, pH 9, 10 mM
      1. Added 0.608g of Tris to distilled water in a 500mL volumetric flask
      2. Mixed for several minutes with a stirbar
      3. Added 1M HCl to obtain a final pH of 9.036
      4. Final conductivity was 172.6 mS
    2. Phosphate buffer: 500 mL, pH 7, 10mM
      1. Added 0.399 monobasic sodium phosphate and 0.369 g dibasic sodium phosphate to distilled water in a 500mL volumetric flask
      2. Added NaOH to reach final pH of 7.058
      3. Final conductivity was 1316 μS.


Matt Hartings How did you make your buffers? What were the final measurements (conductivity and pH) that you took? Did you have to add HCl/NaOH to either both? What were the calculations/measurements that you used for the phosphate.