scholarly journals Komparasi Metode Isolasi DNA Patogen Antraknosa dan Bulai untuk Deteksi PCR

2016 ◽  
Vol 12 (4) ◽  
pp. 124
Author(s):  
Ade Syahputra ◽  
Kikin Hamzah Mutaqin ◽  
Tri Asmira Damayanti
Keyword(s):  
Pure Culture ◽  
Dna Isolation ◽  
Total Yield ◽  
Dna Amplification ◽  
Colletotrichum Acutatum ◽  
Isolation Method ◽  
Peronosclerospora Sorghi ◽  
Dna Purity ◽  
Dna Template ◽  
Commercial Kit

Polymerase chain reaction (PCR) is an important tool for detection, identification and monitoring of quarantine pests in Indonesia. DNA isolation method from target organism is an important step to provide adequate DNA template for performing PCR. Objective of the research was to compare conventional, commercial kit, FTA-card and its modification methods of DNA isolation to be used in PCR detection for Colletotrichum acutatum and Peronosclerospora sorghi from chili and maize, respectively. DNA obtained from various isolation methods were measured using UV-vis nanodrop-spectrophotometry.  DNA amplification was performed using DNA concentration of 15 ng µL-1 from each isolation method with gradual primer concentrations of 0.4; 0.6; 0.8; and 1.0 mM. The highest concentration of DNA was achieved with conventional methods for C. acutatum from pure culture and P. sorghi from maize leaf. Best DNA purity was obtained from isolation method using commercial kit for C. acutatum from infected fruit (1.94) and from conventional method for C. acutatum from pure culture (1.91). The highest total yield of isolated DNA was achieved by modified FTA-card for C. acutatum from pure culture. In general DNA amplification using various primer concentration gave positive results although DNA bands intensity was varied from faint to very bright.  Furthermore PCR optimization using the best primer concentration from previous reaction showed that all DNA templates resulted in thick and bright DNA bands.

scholarly journals Genetic of Salak Pondoh, Gading Varieties and Its Hybrids Based on RAPD Markers

2021 ◽  
Vol 1 (1) ◽  
pp. 5
Author(s):  
Nandariyah Nandariyah ◽  
Parjanto Parjanto ◽  
Pinaka Pinasti Ratu
Keyword(s):  
Molecular Marker ◽  
Rapd Markers ◽  
Dna Isolation ◽  
Dna Amplification ◽  
Isolation Method ◽  
Isolation Methods

<p>A molecular marker of parent and offspring is used to find fast and accurate markers influenced by DNA isolation and amplification. This research aims to find the most suitable DNA isolation and DNA  amplification methods. This study used four DNA isolation methods; namely IM01, IM02, IM03, and IM04. DNA amplification used ten protocols (AP01, AP02, AP03, AP04, AP05, AP06, AP07, AP08, AP09, and AP010). The results of the research showed that the most suitable DNA isolation method for salak was  IM0, and the most suitable DNA amplification for salak was AP04 that produces the highest value of DNA bands.</p><p><strong> </strong></p><p>Keywords: DNA isolation; DNA amplification; hybrids.</p>

scholarly journals Biased nicking of mitochondrial DNA during extraction can be avoided by choice of isolation method

2021 ◽  
Author(s):  
Bruno Marçal Repolês ◽  
Choco Michael Gorospe ◽  
Phong Tran ◽  
Anna Karin Nilsson ◽  
Paulina H. Wanrooij
Keyword(s):  
Skeletal Muscle ◽  
Mitochondrial Dna ◽  
Dna Isolation ◽  
Isolation Procedure ◽  
Isolation Method ◽  
Strand Breaks ◽  
Careful Choice ◽  
Isolation Methods ◽  
Solid Tissues ◽  
Long Range Pcr

AbstractThe integrity of mitochondrial DNA (mtDNA) isolated from solid tissues is critical for analyses such as long-range PCR. We show that a commonly-used DNA isolation procedure preferentially introduces strand breaks into the mtDNA extracted from the skeletal muscle of aged mice, while mtDNA from adult animals is less affected. We present a comparison of mtDNA isolation methods and identify one that avoids this biased loss of muscle mtDNA integrity. Our results highlight the importance of a careful choice of mtDNA isolation method and serve as a resource to researchers planning analysis of mtDNA isolated from solid tissues.

scholarly journals Method Comparison of DNA Isolation and Quantification for Fish and Seafood Authenticity Determination

2018 ◽  
Vol 13 (3) ◽  
pp. 115
Author(s):  
Dwiyitno Dwiyitno ◽  
Stefan Hoffman ◽  
Koen Parmentier ◽  
Chris Van Keer
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Dna Isolation ◽  
Blue Mussel ◽  
Pcr Amplification ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Seafood Products ◽  
Polymerase Chain ◽  
Commercial Kit

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen

scholarly journals Detection of Porcine DNA in Processed Beef Products Using Real Time – Polymerase Chain Reaction

2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Triayu Septiani
Keyword(s):  
Pcr Analysis ◽  
Rt Pcr ◽  
Fresh Meat ◽  
Beef Products ◽  
Dna Purity ◽  
Halal Food ◽  
Porcine Gene ◽  
Amplification Technique ◽  
Processed Products ◽  
Dna Template

Meat is one of food materials which has protein source and mostlyconsumed by non-vegetarian. Consuming halal food is an obligation for every Muslim. Meat processed products usually contaminated by pork. One of technique that is often chosen as an authentication process for proofing halalness of the product is PCR technique, one of PCR technique which most commonly used is RT-PCR. RT-PCR technique was chosen as identification method because it has high accuration for detection of porcine DNA in fresh meat and processed products. RT-PCR is the amplification technique in the specific regions that are restricted by two oligonucleotide with the help of polymerase enzymes. Annealing is the first process of RT-PCR analysis who was primary attachment to the DNA template that determines the specificity and amount of DNA produced. In this study, extraction kit and detection kit were used for analysis Porcine DNA in meatballs. The results obtained from this study were from whole DNA samples, which had DNA purity ranging from 1.82 to 1.93. From the all samples three of them containing porcine DNA. The positive samples shown from amplification curves who was specifically formed when probes reacts with porcine gene.

Methods for Detection of Anticarsia gemmatalis Nucleopolyhedrovirus DNA in Soil

1999 ◽  
Vol 65 (6) ◽  
pp. 2307-2311 ◽  
Author(s):  
R. R. de Moraes ◽  
J. E. Maruniak ◽  
J. E. Funderburk
Keyword(s):  
Laboratory Experiments ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Soil Samples ◽  
Anticarsia Gemmatalis ◽  
Isolation Method ◽  
Viral Dna ◽  
Pcr Inhibitors ◽  
Magnetic Capture ◽  
Pcr Products

ABSTRACT Two methods, phenol-ether and magnetic capture-hybridization (MCH), were developed and compared with regard to their sensitivities and abilities to extract the DNA of the insect baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) from soil and to produce DNA amplifiable by PCR. Laboratory experiments were performed with 0.25 g of autoclaved soil inoculated with different viral concentrations to optimize both methods of baculovirus DNA extraction and to determine their sensitivities. Both procedures produced amplifiable DNA; however, the MCH method was 100-fold more sensitive than the phenol-ether procedure. The removal of PCR inhibitors from the soil appeared to be complete when MCH was used as the viral DNA isolation method, because undiluted aliquots of the DNA preparations could be amplified by PCR. The phenol-ether procedure probably did not completely remove PCR inhibitors from the soil, since PCR products were observed only when the AgMNPV DNA preparations were diluted 10- or 100-fold. AgMNPV DNA was detected in field-collected soil samples from 15 to 180 days after virus application when the MCH procedure to isolate DNA was coupled with PCR amplification of the polyhedrin region.

DNA isolation method for high polysaccharide Lesquerella species

1999 ◽  
Vol 9 (2) ◽  
pp. 111-114 ◽  
Author(s):  
Benjamin Kaufman ◽  
Stacy Richards ◽  
David A Dierig
Keyword(s):  
Dna Isolation ◽  
Isolation Method

High-throughput DNA isolation method for detection of Xylella fastidiosa in plant and insect samples

2011 ◽  
Vol 86 (3) ◽  
pp. 310-312 ◽  
Author(s):  
Jeff A. Brady ◽  
Jennifer B. Faske ◽  
Jessica M. Castañeda-Gill ◽  
Jonathan L. King ◽  
Forrest L. Mitchell
Keyword(s):  
High Throughput ◽  
Dna Isolation ◽  
Xylella Fastidiosa ◽  
Isolation Method
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