Pelling:Protocols/Extracting DNA from Filter Paper

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="930"
   Lab Supplies=Pelling:Lab Supplies
   Lab Safety and MSDS's=Pelling:Lab Safety


Plasmids are often shipped from one lab to another on filter paper. When receiving a plasmid from another lab you will typically get a piece of filter paper with a circle drawn on it in pencil. Usually 1-2μL of plasmid DNA is deposited inside the circle, to extract the DNA:

  • Cut out the circle containing the DNA (do not include the pencil marks).
  • Place in a micro-centrifuge tube with 100μL of 10 mM Tris-HCl, 1 mM EDTA, pH 8.0 buffer (TE Buffer). You can also use dH2O or 10mM Tris buffer.
  • Briefly vortex and let stand for 15min (or overnight in fridge).
  • Use 1-10μL (usually 2μL is good) to transform competent E. coli.
  • Keep tube in the fridge until you are sure that the DNA has been isolated and you are happy with the transformation, transfection and expression of the plasmid.