Dual color fluorescence quantitative detection of specific single-stranded DNA with molecular beacons and nucleic acid dye SYBR Green I

The Analyst ◽  
2012 ◽  
Vol 137 (16) ◽  
pp. 3787 ◽  
Author(s):  
Dong-Shan Xiang ◽  
Guo-Hua Zhou ◽  
Ming Luo ◽  
Xing-Hu Ji ◽  
Zhi-Ke He
Keyword(s):  
Nucleic Acid ◽  
Sybr Green ◽  
Molecular Beacons ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Acid Dye ◽  
Single Stranded Dna ◽  
Dual Color

A label-free signal amplification assay for DNA detection based on exonuclease III and nucleic acid dye SYBR Green I

Talanta ◽  
2013 ◽  
Vol 114 ◽  
pp. 49-53 ◽  
Author(s):  
Aihua Zheng ◽  
Ming Luo ◽  
Dongshan Xiang ◽  
Xia Xiang ◽  
Xinghu Ji ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Signal Amplification ◽  
Dna Detection ◽  
Sybr Green ◽  
Sybr Green I ◽  
Label Free ◽  
Acid Dye ◽  
Exonuclease Iii ◽  
Amplification Assay

Detection of hepatitis B virus with a non-labeled molecular beacon and a nucleic acid dye SYBR Green I

2014 ◽  
Vol 44 (12) ◽  
pp. 1996-2003
Author(s):  
Kun ZHAI ◽  
DongShan XIANG ◽  
LianZhi WANG ◽  
BoAn SHI
Keyword(s):  
Hepatitis B Virus ◽  
Nucleic Acid ◽  
Hepatitis B ◽  
Molecular Beacon ◽  
Sybr Green ◽  
Sybr Green I ◽  
Acid Dye ◽  
B Virus

Fast and sensitive quantitative detection of HIV DNA in whole blood leucocytes by SYBR green I real-time PCR assay

2007 ◽  
Vol 21 (5-6) ◽  
pp. 368-378 ◽  
Author(s):  
Anna Casabianca ◽  
Caterina Gori ◽  
Chiara Orlandi ◽  
Federica Forbici ◽  
Carlo Federico Perno ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Whole Blood ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Pcr Assay ◽  
Hiv Dna

scholarly journals Quantitative Detection of Listeria monocytogenes in Biofilms by Real-Time PCR

2005 ◽  
Vol 71 (4) ◽  
pp. 2190-2194 ◽  
Author(s):  
Morgan Guilbaud ◽  
Pierre de Coppet ◽  
Fabrice Bourion ◽  
Cinta Rachman ◽  
Hervé Prévost ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Method ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Pcr Assay

ABSTRACT A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 × 102 CFU/cm2.

Real-time PCR assay for sensitive organ detection and epidemic investigation of Turbot reddish body iridovirus

2009 ◽  
Vol 6 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Wu Cheng-Long ◽  
Shi Cheng-Yin ◽  
Huang Jie ◽  
Kong Xiao-Yu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Viral Genome ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Linear Coefficient ◽  
Sensitive Organ

AbstractA rapid and sensitive real-time polymerase chain reaction (PCR) assay coupled with SYBR Green I chemistry was developed for the quantitative detection of Turbot reddish body iridovirus (TRBIV) isolated from farmed turbot (Scophthalmus maximus). A 152 bp DNA fragment from the TRBIV major capsid protein (MCP) gene was involved in the real-time PCR (RT-PCR) assay using the Roter Gene 3000 sequence detection system. The PCR mixture contained a fluorescent dye, SYBR Green I, which exhibited fluorescence enhancement when bound to double-stranded (ds) DNA. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. The positive control plasmid, pUCm-T/TRBIV MCP, containing the target sequence, was quantified to make a standard curve for sample detection after serial tenfold dilution. Linear coefficient correlations between the cycle threshold (CT) value and logarithmic positive plasmid concentration were close to one (R2=0.9952) and the detection limit of the assay was 102 copies of positive plasmids. The quantitative detection of virus in different tissues from TRBIV-infected fish showed that the spleen and kidney contained the largest number of viral particles (5.23×106 and 2.18×106 viral genome copies/mg tissue, respectively), while no viral DNA was detected in the muscular tissue. The molecular epidemic investigation of TRBIV showed that many cultured turbots were infected and TRBIV has become epidemic in turbot farms located along the Shandong peninsula. The virus number varied from 1.27×102 to 2.33×106 viral genome copies/mg tissue in spleens of infected turbot. These results suggest that the RT-PCR assay reported here can be used as a rapid, sensitive and quantitative method for TRBIV.

scholarly journals Sensitive Determination of Microbial Growth by Nucleic Acid Staining in Aqueous Suspension

2006 ◽  
Vol 72 (1) ◽  
pp. 87-95 ◽  
Author(s):  
Willm Martens-Habbena ◽  
Henrik Sass
Keyword(s):  
Nucleic Acid ◽  
Sybr Green ◽  
Sybr Green I ◽  
Epifluorescence Microscopy ◽  
Most Probable Number ◽  
Staining Procedure ◽  
Fluorescence Measurement ◽  
Probable Number ◽  
Microbial Cells

ABSTRACT The determination of cell numbers or biomass in laboratory cultures or environmental samples is usually based on turbidity measurements, viable counts, biochemical determinations (e.g., protein and lipid measurements), microscopic counting, or recently, flow cytometric analysis. In the present study, we developed a novel procedure for the sensitive quantification of microbial cells in cultures and most-probable-number series. The assay combines fluorescent nucleic acid staining and subsequent fluorescence measurement in suspension. Six different fluorescent dyes (acridine orange, DAPI [4′,6′-diamidino-2-phenylindole], ethidium bromide, PicoGreen, and SYBR green I and II) were evaluated. SYBR green I was found to be the most sensitive dye and allowed the quantification of 50,000 to up to 1.5 × 108 Escherichia coli cells per ml sample. The rapid staining procedure was robust against interference from rRNA, sample fixation by the addition of glutaric dialdehyde, and reducing agents such as sodium dithionite, sodium sulfide, and ferrous sulfide. It worked well with phylogenetically distant bacterial and archaeal strains. Excellent agreement with optical density measurements of cell increases was achieved during growth experiments performed with aerobic and sulfate-reducing bacteria. The assay offers a time-saving, more sensitive alternative to epifluorescence microscopy analysis of most-probable-number dilution series. This method simplifies the quantification of microbial cells in pure cultures as well as enrichments and is particularly suited for low cell densities.

Sign in / Sign up

Export Citation Format

Share Document