Purification of PCR products by PEG-precipitation

  • mix equal volumes of PCR and PEG solution (vortex!)
  • incubate 10 min at room temperature
  • centrifuge 10 min at 12 000 - 14 000 rpm
  • remove the supernatant
  • add 100µl 96% ethanol
  • centrifuge 10 min at 12 000 - 14 000 rpm
  • remove the supernatant
  • air dry 5 min at room temperature
  • resolve in a suitable volume of 10 mM TRIS/Cl pH 7.5 - 8

for 1000mL PEG-solution:

49.2g
1.2g
262.0g

NaOAc
MgCl
2 x 6H2O
PEG 8000

Ref:

Rosenthal A, Coutelle O, Craxton M. Large-scale production of DNA sequencing templates by microtitre format PCR. Nucleic Acids Res. 1993 Jan 11;21(1):173-4 [Pubmed]