A streamlined protocol for emulsion polymerase chain reaction and subsequent purification.
Compartmentalization of polymerase chain reaction (PCR) reduces artifacts, especially when complex libraries are amplified. It allows clonal amplification of templates from complex mixtures in a bias-free manner. Here we describe a rapid, straightforward, and easy protocol for PCR in a water-in-oil emulsion (ePCR) including sample recovery by DNA purification. Furthermore, no special laboratory equipment is needed and inexpensive components are used. Therefore, our flexible protocol allows ePCR to be readily implemented in daily routine experiments for a broad range of applications.
Department of Vertebrate Genomics, Max Planck Institute for Molecular Genetics, Ihnestrasse 63-73, 14195 Berlin, Germany.
Tatjana
T
Schütze
Florian
F
Rubelt
Julia
J
Repkow
Nicole
N
Greiner
Volker A
VA
Erdmann
Hans
H
Lehrach
Zoltán
Z
Konthur
Jörn
J
Glökler
D005591:Chemical Fractionation / Q000379:methods*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D004655:Emulsions; D006801:Humans; D009821:Oils / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors; D014867:Water / Q000737:chemistry*
D004655:Emulsions; D009821:Oils; D014867:Water; D004247:DNA