scholarly journals Quantitative real-time PCR method to detect changes in specific transcript and total RNA amounts

2004 ◽  
Vol 7 (1) ◽  
Author(s):  
Daniel Z. Skinner ◽  
Kwang-Hyun Baek
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Total Rna ◽  
Specific Transcript ◽  
Pcr Method

scholarly journals Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7188
Author(s):  
Long Yu ◽  
Xiaofei Wu ◽  
Yang Yu ◽  
Limei Shi ◽  
Min Zhang
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Lake Taihu ◽  
Rna Extraction ◽  
Sybr Green ◽  
Quantitative Detection ◽  
Control Group ◽  
Quantitative Real Time Pcr ◽  
Total Rna ◽  
Pcr Method

In this study, a SYBR Green quantitative real-time PCR method was established and applied. Relative expression of the synthetic genes from Microcystis gas vesicles (gvpC), algal toxin genes (mcyA), and polysaccharides (espL) from water and sediments of Meiliang Bay and from the center of Lake Taihu were tested from January to June, 2017. Indoor Microcystis aeruginosa was used as the control group. The kit for total RNA extraction in Microcystis was optimized. Results showed that the optimized kit extracted high-concentrations and high-quality total RNA from Microcystis. The extraction purity and concentration were significantly higher than those extracted by the original kit. The transcription level of gvpC increased gradually until a peak was reached in March. However, expression of gvpC decreased continuously at the proliferating and floating stages of Cyanobacterial biomass. The maximum level of expression of gvpC in April in comparison to expression of mcyA in March occurred first. We found that the SYBR Green qRT-PCR method, which is characterized by high specificity, repeatability, is rapid, and can be used for quantitative detection of expression of gvpC, mcyA, and espL. The recruitment of cyanobacteria is the process in which cyanobacteria in the sediment began to regain their activity, started to grow and migrated to the water column.

Collaborative Validation of an Event-Specific Quantitative Real-Time PCR Method for Genetically Modified Rice Event TT51-1 Detection

2013 ◽  
Vol 61 (25) ◽  
pp. 5953-5960 ◽  
Author(s):  
Yuhua Wu ◽  
Litao Yang ◽  
Yinglong Cao ◽  
Guiwen Song ◽  
Ping Shen ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified ◽  
Quantitative Real Time Pcr ◽  
Genetically Modified Rice ◽  
Pcr Method

A Quantitative Real-Time PCR Method Using an X-Linked Gene for Sex Typing in Pigs

2012 ◽  
Vol 54 (2) ◽  
pp. 493-496 ◽  
Author(s):  
Maria Ballester ◽  
Anna Castelló ◽  
Yuliaxis Ramayo-Caldas ◽  
Josep M. Folch
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Sex Typing ◽  
Linked Gene ◽  
Pcr Method

Quantitative Detection of Campylobacter jejuni on Fresh Chicken Carcasses by Real-Time PCR

2007 ◽  
Vol 70 (6) ◽  
pp. 1373-1378 ◽  
Author(s):  
ANNA-CLARA RÖNNER ◽  
HANS LINDMARK
Keyword(s):  
Detection Limit ◽  
Real Time ◽  
Dna Extraction ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Close Correlation ◽  
Quantitative Detection ◽  
Quantitative Real Time Pcr ◽  
Direct Plating ◽  
Pcr Method

Campylobacter jejuni infection is a significant cause of foodborne gastroenteritis worldwide. Consumption and handling of poultry products is believed to be the primary risk factor for campylobacteriosis. Risk assessments require quantitative data, and C. jejuni is enumerated usually by direct plating, which sometimes allows growth of non-Campylobacter bacteria. The objective of the present study was to develop a quantitative real-time PCR method (q-PCR) for enumerating C. jejuni in chicken rinse without a culturing step. The procedure to obtain the template for the PCR assay involved (i) filtration of 10 ml of chicken rinse, (ii) centrifugation of the sample, and (iii) total DNA extraction from the pellet obtained using a commercial DNA extraction kit. The detection limit of the method was comparable to that for plating 100 μl of chicken rinse on modified charcoal cefoperazone deoxycholate agar, and the detection limit could be further improved 10-fold by concentrating the DNA eluate by ethanol precipitation. A close correlation for spiked chicken rinse was obtained for the results of the quantitative real-time PCR method and direct plating (r = 0.99). The coefficient of correlation for the methods was 0.87 when samples from chicken carcasses on the slaughter line were analyzed, whereas a lower correlation (r = 0.76) was obtained when samples from retail carcasses were analyzed. Greater variation in the proportion of dead and/or viable but not culturable Campylobacter types in the retail samples may explain the decreased correlation between the methods. Overall, the new method is simple and fast and the results obtained are closely correlated with those for direct plating for samples containing a low proportion of dead Campylobacter cells.

scholarly journals A quantitative real-time PCR method for absolute telomere length

BioTechniques ◽  
2008 ◽  
Vol 44 (6) ◽  
pp. 807-809 ◽  
Author(s):  
Nathan J. O'Callaghan ◽  
Varinderpal S. Dhillon ◽  
Philip Thomas ◽  
Michael Fenech
Keyword(s):  
Telomere Length ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Pcr Method

scholarly journals A human fecal contamination score for ranking recreational sites using the HF183/BacR287 quantitative real-time PCR method

2018 ◽  
Vol 128 ◽  
pp. 148-156 ◽  
Author(s):  
Yiping Cao ◽  
Mano Sivaganesan ◽  
Catherine A. Kelty ◽  
Dan Wang ◽  
Alexandria B. Boehm ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fecal Contamination ◽  
Quantitative Real Time Pcr ◽  
Pcr Method

Analysis of alternative promoter usage in expression of HSD11B1 including the development of a transcript-specific quantitative real-time PCR method

2011 ◽  
Vol 191 (1-3) ◽  
pp. 104-112 ◽  
Author(s):  
Claudia A. Staab ◽  
Jochen P. Stegk ◽  
Sierk Haenisch ◽  
Elisabeth Neiß ◽  
Katrin Köbsch ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Alternative Promoter ◽  
Quantitative Real Time Pcr ◽  
Pcr Method ◽  
Promoter Usage

Analysis of community structures in anaerobic processes using a quantitative real-time PCR method

2005 ◽  
Vol 52 (1-2) ◽  
pp. 85-91 ◽  
Author(s):  
Y. Yu ◽  
C. Lee ◽  
S. Hwang
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rrna Gene ◽  
Quantitative Real Time Pcr ◽  
Community Structures ◽  
Specific Primer ◽  
Anaerobic Processes ◽  
Promising Tool ◽  
Pcr Method ◽  
Group Specific Primer

The methanogenic community structures of four different anaerobic processes were characterized using a quantitative real-time PCR with group-specific primer and probe sets targeting the 16S rRNA gene (rDNA). The group specific primer and probe sets were developed and used to detect the orders Methanosarcinales, and the families Methanosarcinaceae and Methanosaetaceae. Two separate sets targeting the domains Archaea and Bacteria were also used. Each microbial population in different anaerobic processes was determined and the relative abundance in the system was compared with each other. Dominant methanogenic populations and the community structures in the processes were varied by hydraulic retention time and acetate concentration. This indicates that the real-time PCR method with the primer and probe sets is a promising tool to analyze community structures in anaerobic processes.

scholarly journals A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

2010 ◽  
Vol 51 (6) ◽  
pp. 603-610 ◽  
Author(s):  
B. Llorente ◽  
F. Bravo-Almonacid ◽  
C. Cvitanich ◽  
E. Orlowska ◽  
H.N. Torres ◽  
...  
Keyword(s):  
Phytophthora Infestans ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
In Planta ◽  
Pcr Method
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