1.2 DNA Quantification

1.2 DNA Quantification margaret Wed, 11/16/2011 - 18:07

Goal for this lab:

Quantify Arabidopsis DNA by two methods. (It is hard to finish all the analysis in 3 hours – better if this happens on a Monday 4 hour lab. Otherwise save the gel analysis till the next period, along with the Wiki.)

For pairs of students
make: 1 mL aliquots DB (10 mM Tris pH 7.5) or T10E1
make: 100 µL aliquots 6X loading dye
make: 25 µL aliquots MassRuler High Range

6X Loading dye to be used throughout the semester: 100 µL

MassRuler™ DNA Ladder, High Range Fermentas #SM0393 Give them 25µL in a microfuge tube

Materials for DNA quant

Materials for DNA quant kdorfman Thu, 01/26/2012 - 16:41

EQUIPMENT

  • Turn on the spectrophotometers
  • Cut and distribute diaper pads
  • Make EtBr waste containers
  • Diapered area (to catch ethidium bromide drips)
  • sterile
    • microfuge tubes
    • tips
  • paper towels for handling gels
  • Cuvettes
    • Krackeler 478-759220 pack of 100ultra micro cuvettes, 15 mm window ht, Brand UV $71 list price
    • Fisher 13-878-123 BRAND* UV-Cuvette Disposable Cuvets from BrandTech* Ultramicro; 15mm window; 100/Pk $49.66 available at stockroom

Reading should be below 0.3 to stay in linear range. See graph showing loss of linearity for fluorescein (in 1M NaOH) absorbance in Jenova spec above Abs=0.3