BMBMethod:EtOHDNA

From OpenWetWare
Jump to: navigation, search

<owwmenu font="arial, helvetica, sans-serif" bold="1" color="black" bgcolor="white" hovercolor="black" bghovercolor="gray" topFontSize="10" fontSize="8" pagewidth="650" image="OldMain2.jpg" lab="BMBMethod"> Home= Meetings=#,Schedule=Schedule, Past=Past, Presentations=Presentations Protocols=Protocols, Protein=Protocols#Protein, DNA=Protocols#DNA, RNA=Protocols#RNA, Other=Protocols#Other Tips & Tricks=TipsandTricks, Protein=TipsandTricks#Protein, DNA=TipsandTricks#DNA, RNA=TipsandTricks#RNA, Other=TipsandTricks#Other Equipment=Equipment, Departmental=Equipment#BMBDepartment, Personal=Equipment#Personal, Other=Equipment#Other Reagents=Reagents, Departmental=Reagents#Departmental, Cheap Buys=Reagents#Cheap Reagents </owwmenu>

EtOH Precipitation of DNA

This protocol can be used to concentrate DNA, change buffers, or get rid of interfering materials before restriction enzyme digestion, ligation, or whatever.

  • Add 1/10 volume of 3M sodium acetate (pH 5.2) to DNA solution
  • Add 2.5 volumes of 100% ethanol
  • Vortex, store tube at -80°C for 15 min
  • Centrifuge the tube to pellet DNA at 14,000 rpm for 15 min
  • Carefully pour off the supernatant, taking care not to disturb the pellet
  • Add 150ul 70% ethanol to wash the pellet
  • Centrifuge at 14,000 rpm for 5min
  • Discard the supernatant
  • Spin down to collect any residual supernatant, then pipette it off
  • Dry the pellet in air or in SpeedVac
  • DNA pellet can be re-dissolved in ddH2O or TE buffer.