Pre-PCR Processing, PCR Facilitators and Stabilizer Additives

Pre-PCR processing of samples.

Pre-PCR processing: strategies to generate PCR-compatible samples.

Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed t...

Improved blood sample processing for PCR.

To simplify our procedure for blood sample processing for PCR, we introduced a simpler, shorter, and more cost-effective method for the separation of peripheral blood lymphocytes (PBL) and DNA extraction for amplification with Taq polymerase. By this method, blood samples are processed in two simple 15-min steps: (1) separation of PBLs from whole blood by red blood cell lysis with the Roche Spe...

Evaluation of pre-PCR processing approaches for enumeration of Salmonella enterica in naturally contaminated animal feed.

AIMS Three pre-PCR processing strategies for the detection and/or quantification of Salmonella in naturally contaminated soya bean meal were evaluated. METHODS AND RESULTS Methods included: (i) flotation-qPCR [enumeration of intact Salmonella cells prior to quantitative PCR (qPCR)], (ii) MPN-PCR (modified most probable number method combined with qPCR) and (iii) qualitative culture enrichment...

Expression-PCR (E-PCR): overview and applications.

1Department of Immunology, Waiter Reed Army Institute of Research, Washington, D.C. 20307-5100; 2Tropical Disease Unit, Division of Infectious Diseases, The Toronto Hospital and The University of Toronto, Toronto, Ontario, Canada M5G 2C4 In vitro transcription and translation are powerful tools for examining the s t ructure-funct ion relationships of proteins. Genes can be cloned into plasmid v...