Use of citric acid for neutralizing polymerase chain reaction inhibition by chlorogenic acid in potato extracts.
Polyphenolics are a widely distributed class of plant constituents implicated in the inhibition of the polymerase chain reaction (PCR). Since chlorogenic acid forms the bulk of phenolics in the potato tuber, a range of chlorogenic acid concentrations were tested for inhibition of reverse transcription (RT) and PCR steps. The RT step was inhibited at chlorogenic acid concentrations of 6.0 to 7.5 microg, while the PCR step was inhibited at much lower concentrations of 1.2 to 1.8 microg. Incorporation of 1.2% citric acid at the nucleic acid extraction step prevented darkening of the extract and neutralized the inhibitory effect of chlorogenic acid.
Potato Research Centre, Agriculture and Agri-Food Canada, Fredericton, NB. singhr@em.agr.ca
R P
RP
Singh
M
M
Singh
R R
RR
King
D002726:Chlorogenic Acid / Q000494:pharmacology*; D019343:Citric Acid / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D010936:Plant Extracts / Q000737:chemistry; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000187:drug effects*; D011198:Solanum tuberosum / Q000737:chemistry / Q000821:virology*
D010936:Plant Extracts; D012367:RNA, Viral; D019343:Citric Acid; D002726:Chlorogenic Acid