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).

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 Detection of Acanthamoeba spp. in two major water reservoirs in the Philippines

2020 ◽  
Vol 18 (2) ◽  
pp. 118-126 ◽  
Author(s):  
Giovanni Milanez ◽  
Frederick Masangkay ◽  
Frieda Hapan ◽  
Thea Bencito ◽  
Marcus Lopez ◽  
...  
Keyword(s):  
Water Samples ◽  
Gel Electrophoresis ◽  
Water Security ◽  
The Philippines ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Water Reservoirs ◽  
Chain Reaction ◽  
Pcr Products ◽  
Polymerase Chain

Abstract Water reservoirs are important manmade structures providing water security to deliver clean and safe water for drinking and other purposes to the community. Eighty water samples were collected from Magat and Ipo water reservoirs using purposive sampling between November 2018 and January 2019. Water samples were collected in sterile containers for testing. The samples were cultured in non-nutrient agar and lawned with Escherichia coli and incubated at 33 °C. Twelve out of the 80 (15%) water samples were positive for amoebic growth. Light and scanning electron microscopy (SEM) revealed double-walled cystic stages and were initially identified as Acanthamoeba spp. based on morphological characteristic in reference to Page's established criteria. Their extracted DNAs were used in polymerase chain reaction using JDP1 and JDP2 primers and confirmed the presence of Acanthamoeba DNA in agarose gel electrophoresis. Aligned sequences from PCR products were deposited in GenBank under accession numbers MK886460, MK909919, MK905437, MK910997, MK911021 and MK886514. The presence of potentially pathogenic Acanthamoeba spp. in water reservoirs is considered a potential risk for public health, requiring appropriate processing of water in treatment plants.

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 High Resolution Microsatellite Marker Analysis of Some Rice Landraces Using Metaphor Agarose Gel Electrophoresis

2016 ◽  
Vol 20 (1) ◽  
pp. 54 ◽  
Author(s):  
K. Kristamtini ◽  
T. Taryono ◽  
Panjisakti Basunanda ◽  
Rudi Hari Murti
Keyword(s):  
Microsatellite Markers ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Molecular Techniques ◽  
Agarose Gel ◽  
Chain Reaction ◽  
Rice Landraces ◽  
Polymorphic Pattern ◽  
Polymerase Chain ◽  
Microsatellite Marker Analysis

Microsatellite markers or simple sequences repeats are DNA - based molecular techniques that areused to see the different among accessions and inbred lines. There are three methods to analysis the results ofthe polymerase chain reaction of microsatellite markers namely polyacrylamide gel electrophoresis (PAGE),capillary electroforesis, and Metaphor Agarose Gel Electroforesis (MAGE), and the Use of MAGE assessedmore easily and economically the polymorphic pattern of DNA markers. This study aimed to obtain fast,effective and efficient in term of easy and cheap technique to identify microsatellite markers of some blackrice cultivars and F2 populations from crosses between black with white rice. The results showed that MAGEsuccessfully separated clearly SSRs alleles with different sizes of less than 25 bp .

scholarly journals Rapid Detection and Quantification of Rhynchosporium secalis in Barley Using a Polymerase Chain Reaction

2011 ◽  
Vol 42 (No. 3) ◽  
pp. 111-114
Author(s):  
J. Gubiš ◽  
M. Hudcovicová ◽  
M. Gubišová
Keyword(s):  
Gel Electrophoresis ◽  
Pcr Primers ◽  
Rhynchosporium Secalis ◽  
Agarose Gel Electrophoresis ◽  
Artificial Infection ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Green Approach ◽  
Total Dna ◽  
Polymerase Chain

PCR primers for diagnosis of Rhynchosporium secalis in seed samples of barley were developed. For the quantification of the pathogen in seed samples a real-time PCR with SYBR Green approach was used. Amounts from 1.8 to 419.1 pg of R. secalis DNA per 100 ng of total DNA were detected in 18 samples of barley seeds contaminated by R. secalis in field conditions. The correctness of this quantitative analysis was checked using an artificial infection of seeds with 1, 2, 5 and 20% level of infection by R. secalis. The level of contamination of artificially infected samples decreased with a lowering amount of added seed powder contaminated by the pathogen, the correlation coefficient for this analysis was 0.98. While the primer pair used in these analyses shows cross-reactions with other pathogens (P. teres, Drechslera tritici-repentis, F. culmorum and F. poe), it is recommended to check the products of RT-PCR by agarose-gel electrophoresis, in which these pathogens are easily distinguishable from R. secalis by different lengths of the amplified fragments.  

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