scholarly journals Improved quantitative real-time PCR assays for enumeration of harmful algal species in field samples using an exogenous DNA reference standard

2005 ◽  
Vol 3 (9) ◽  
pp. 381-391 ◽  
Author(s):  
Kathryn J. Coyne ◽  
Sara M. Handy ◽  
Elif Demir ◽  
Edward B. Whereat ◽  
David A. Hutchins ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Algal Species ◽  
Reference Standard ◽  
Quantitative Real Time Pcr ◽  
Exogenous Dna ◽  
Field Samples ◽  
Pcr Assays

scholarly journals Identification and Validation of Suitable Housekeeping Genes for Normalizing Quantitative Real-Time PCR Assays in Injured Peripheral Nerves

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e105601 ◽  
Author(s):  
Giovanna Gambarotta ◽  
Giulia Ronchi ◽  
Olivier Friard ◽  
Pantaleo Galletta ◽  
Isabelle Perroteau ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Nerves ◽  
Housekeeping Genes ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays

Quantitative real-time PCR assays to detect DNA degradation in soy-based food products

2009 ◽  
Vol 89 (7) ◽  
pp. 1137-1144 ◽  
Author(s):  
Sarah R Murray ◽  
Ruth C Butler ◽  
Gail M Timmerman-Vaughan
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Food Products ◽  
Dna Degradation ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays

scholarly journals Development of Quantitative Real-Time PCR Assays for Detection and Quantification of Surrogate Biological Warfare Agents in Building Debris and Leachate

2007 ◽  
Vol 73 (20) ◽  
pp. 6557-6565 ◽  
Author(s):  
Pascal E. Saikaly ◽  
Morton A. Barlaz ◽  
Francis L. de los Reyes
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genomic Dna ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Fate And Transport ◽  
Biological Warfare ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays ◽  
Detection And Quantification

ABSTRACT Evaluation of the fate and transport of biological warfare (BW) agents in landfills requires the development of specific and sensitive detection assays. The objective of the current study was to develop and validate SYBR green quantitative real-time PCR (Q-PCR) assays for the specific detection and quantification of surrogate BW agents in synthetic building debris (SBD) and leachate. Bacillus atrophaeus (vegetative cells and spores) and Serratia marcescens were used as surrogates for Bacillus anthracis (anthrax) and Yersinia pestis (plague), respectively. The targets for SYBR green Q-PCR assays were the 16S-23S rRNA intergenic transcribed spacer (ITS) region and recA gene for B. atrophaeus and the gyrB, wzm, and recA genes for S. marcescens. All assays showed high specificity when tested against 5 ng of closely related Bacillus and Serratia nontarget DNA from 21 organisms. Several spore lysis methods that include a combination of one or more of freeze-thaw cycles, chemical lysis, hot detergent treatment, bead beat homogenization, and sonication were evaluated. All methods tested showed similar threshold cycle values. The limit of detection of the developed Q-PCR assays was determined using DNA extracted from a pure bacterial culture and DNA extracted from sterile water, leachate, and SBD samples spiked with increasing quantities of surrogates. The limit of detection for B. atrophaeus genomic DNA using the ITS and B. atrophaeus recA Q-PCR assays was 7.5 fg per PCR. The limits of detection of S. marcescens genomic DNA using the gyrB, wzm, and S. marcescens recA Q-PCR assays were 7.5 fg, 75 fg, and 7.5 fg per PCR, respectively. Quantification of B. atrophaeus vegetative cells and spores was linear (R 2 > 0.98) over a 7-log-unit dynamic range down to 101 B. atrophaeus cells or spores. Quantification of S. marcescens (R 2 > 0.98) was linear over a 6-log-unit dynamic range down to 102 S. marcescens cells. The developed Q-PCR assays are highly specific and sensitive and can be used for monitoring the fate and transport of the BW surrogates B. atrophaeus and S. marcescens in building debris and leachate.

Monitoring Lactobacillus plantarum in grass silages with the aid of 16S rDNA-based quantitative real-time PCR assays

2006 ◽  
Vol 29 (1) ◽  
pp. 49-58 ◽  
Author(s):  
Michael Klocke ◽  
Kerstin Mundt ◽  
Christine Idler ◽  
Joseph McEniry ◽  
Padraig O’Kiely ◽  
...  
Keyword(s):  
Real Time ◽  
16S Rdna ◽  
Lactobacillus Plantarum ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays
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