Detection of picornavirus genomic and template RNA strands by a novel semi-nested polymerase chain reaction-technique and agarose gel electrophoresis

1995 ◽  
Vol 16 (1) ◽  
pp. 338-340 ◽  
Author(s):  
John Gow ◽  
Philip Cash ◽  
Wilhelmina Behan ◽  
Frances McGarry ◽  
Kathleen Simpson ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Polymerase Chain Reaction Technique ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

Molecular Beacon Polymerase Chain Reaction Detection of Escherichia coli O157:H7 in Milk

2000 ◽  
Vol 63 (7) ◽  
pp. 855-859 ◽  
Author(s):  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Molecular Beacon ◽  
Skim Milk ◽  
Agarose Gel ◽  
Escherichia Coli O157 ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Polymerase Chain

A fluorescently labeled oligonucleotide probe (molecular beacon) was applied to detect Escherichia coli O157:H7 in artificially contaminated skim milk during polymerase chain reaction (PCR) amplification of extracted DNA. The probe was designed to hybridize with a region of the slt-II gene coding for the A subunit and to fluoresce when the hairpin-stem conformation was linearized upon hybridization to the target sequence. The molecular beacon was incorporated into PCR reactions containing DNA extracted from artificially contaminated skim milk. The degree of fluorescence was monitored in PCR reactions containing 103, 105, and 107 CFU of E. coli O157:H7 per ml and was found to correlate with the amount of template in each reaction. Fluorescence significantly increased above background levels by cycle 8, 14, or 14 in reactions containing DNA from the 107-, 105-, or 103-CFU/ml template, respectively (P < 0.05). Molecular beacon PCR demonstrated positive results more rapidly than traditional agarose gel electrophoresis analysis of PCR products. Use of molecular beacons allows real-time monitoring of PCR reactions, and the closed-tube format allows simultaneous detection and confirmation of target amplicons without the need for agarose gel electrophoresis and/or Southern blotting. This is the first report of a stem-and-loop molecular beacon being applied for direct detection of a pathogen in food.

scholarly journals Polyvinylpyrrolidone-Agarose Gel Electrophoresis Purification of Polymerase Chain Reaction-Amplifiable DNA from Soils

1993 ◽  
Vol 59 (6) ◽  
pp. 1972-1974 ◽  
Author(s):  
Charles C. Young ◽  
Robert L. Burghoff ◽  
Lois G. Keim ◽  
Vera Minak-Bernero ◽  
James R. Lute ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals TECHNICAL SHEET OF COMPARATIVE STUDY OF TWO TECHNIQUES CONTROL OF DNA FROM YEAST ISOLATES PRODUCING PECTINASE

2020 ◽  
Vol 8 (9) ◽  
pp. 979-983
Author(s):  
Coulibaly Wahauwouele Hermann ◽  
◽  
Bouatenin Koffi Maizan Jean-Paul ◽  
Kouame Kohi Alfred ◽  
Amoikon Tiemele Laurent Simon ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Biology ◽  
Spectrophotometric Method ◽  
Gel Electrophoresis ◽  
Extraction Method ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Polymerase Chain

Polymerase Chain Reaction (PCR) is a widely used technique in the field of molecular biology to rapidly make very large number of copies of a specific DNA sample for detailed studies. The success of the technique however is dependent on the on the quality, i.e purity of the extracted DNA specimen. The aim of this study was to evaluate the quality of extracted DNA from pectinase producing yeast to determine the suitability of the extraction method to produce pure extract without non-inhibiting substances.In this study, DNA extracts from six (06) isolates of pectinase-producing yeasts were quantitatively and qualitatively analyzed using NanoDrop spectrophotometry and agarose gel electrophoresis methods. These analyses showed that the concentration of DNA extracts from the isolates evaluated by the NanoDrop spectrophotometric method ranged from 403.8 to 1082.4 ng/µL and the purity index A 260/280 was between 2.03 and 2.11. In sum, agarose gel electrophoresis showed that the intensity of the DNA bands was irregular and not necessarily in line with the data provided by the NanoDrop spectrophotometry.

scholarly journals Sexing as a tool for reinsertion of Amazona aestiva parrots to nature: use of less invasive technique

2021 ◽  
Vol 10 (12) ◽  
pp. e380101220601
Author(s):  
Carine Novato Ramos ◽  
Tácita Borges Barros ◽  
Edma Santos de Antonio ◽  
Bernardo Mirabal ◽  
Márcio Borba da Silva ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Invasive Technique ◽  
Molecular Sexing ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Behavioral Studies ◽  
Sample Composition ◽  
Polymerase Chain ◽  
Primer Sets

The sexing birds is considered an important tool for behavioral studies and programs for the reintroduction of animals into the wild. Several techniques are used for this purpose, such as laparoscopy, magnetic resonance and molecular sexing. The first are considered more invasive and stressful for the animal, and the last is considered the most accurate. According to it, the aim of this study was to compare the effectiveness of using three sets of primers in the molecular sexing process of true parrots (Amazona aestiva). Blood samples from 10 animals were collected at a Wildlife Screening Center (CETAS) in Bahia, Brazil. The DNA was extracted and the molecular markers amplified by Polymerase Chain Reaction (PCR) using primer pairs P2/P8, 1237L/1272H and 2250F/2718R. The amplified material was visualized with electrophoresis performed at 2% agarose and 12.5% polyacrylamide gels. Among the primer sets used, the 2250F/2718R pair showed the best results for the sexing process, including visualization of the amplified products on an agarose gel. Agarose gel electrophoresis is considered to be faster and cheaper. The results revealed a sample composition of 5 males (0.5) and 5 females (0.5).

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