Estimate pBR322 and pGFP(LVA) DNA Concentrations from Prep

 

Materials:

Methods:

1. Set up a blank cuvette, with 0.7 ml Tris-HCl, and blank the spectrophotometer.
2. Set up 1 ml Tris-HCl in each of 2 eppendorf tubes, and label tubes.
3. Aliquot 50   of pBR322 and pGFP(LVA) solutions each into the separate tubes. Mix by inverting tube 4-6 times.
4. Aliquot 0.7 ml solution into cuvette. Measure the for each tube.
5. Compute the DNA concentration, using 1 OD = 50  /ml.

Results: ( 2/5/99)

• For pGFP(LVA), used 50   in 1 ml, and got .

  dilution factor = 20:1

   /ml = 0.08  /ml

  0.08  /ml (dilution factor)

  This is too little, especially the OD reading. Also, analytical gel did not show any presence of this plasmid.

• For pBR322, used 50   in 0.6 ml, and got .

  dilution factor = 12:1

   /ml = 1.5  /ml

  1.5  /ml (dilution factor)

  Analytical gel also showed presence of this plasmid.