Optimisation of PCR reactions using primer chessboarding.
In-house polymerase chain reaction (PCR) assays are now an integral part of the work of most diagnostic microbiological laboratories. Despite the availability of commercial reagent 'master-mixes' of some PCR reagents, the optimisation of primers still poses a significant problem. Here, we describe a simple method to assess the concentration of primer needed in single round, multiplex, nested and 'real-time' PCR procedures.
Gunson
Rory
R
West Of Scotland Specialist Virology Centre, A Member of the UK Clinical Virology Network, Gartnavel General Hospital, 1053 Great Western Road, G12 OYN, Glasgow, UK. w.carman@vir.gla.ac.uk
Gillespie
Graeme
G
F Carman
William
W
D016477:Artifacts; D003199:Computer Systems; D017931:DNA Primers / Q000737:chemistry*; D018076:DNA, Complementary / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D009994:Osmolar Concentration; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D018076:DNA, Complementary