Monthly Archives: July 2016

Ethanol precipitation of DNA

I’ve chosen to take steps to improve the quality of the DNA I extracted over the winter and early spring from the Porites samples I collected in Nov. 2015. Today I selected several samples for which I will plan to generate ddRAD /Epi RAD libraries and subjected them to ethanol precipitation. Here is the protocol I followed (adapted from this):

  • 1/10 volume of 3 M Na-Acetate, pH 5.2 , added to sample
  • 2.2 volumes of ice-cold 100% ethanol added to sample
  • sample mixed and stored at -20°C for 1.5 hours to precipitate the DNA
  • DNA pelleted by centrifugation at full speed (21,000 x g) in microcentrifuge for 30 minutes
  • ethanol oured off and pellet washed twice with room-temperature 70% ethanol
  • DNA pellet air-dried
  • DNA resuspended in 50 μl Qiagen AE buffer

July/August goals

My goals for the remainder of July are to obtain DNA samples of high quality and purity for ddRAD / EpiRad library generation. This includes performing ethanol precipitation of the DNA samples I have.

For August, my goal is to generate the RAD libraries and submit them for sequencing. Maybe I will even get to start in on the bioinformatics.