scholarly journals Comparative Analysis of the Genomic DNA Isolation Methods on Inula sp. (Asteraceae)

2016 ◽  
Vol 8 (4) ◽  
pp. 451-455
Author(s):  
Emre SEVİNDİK ◽  
Fatih COŞKUN ◽  
Zehra Tuğba MURATHAN ◽  
Mehmet Yavuz PAKSOY ◽  
Veysel UZUN
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Low Cost ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Dna Amount ◽  
Genomic Dna Isolation ◽  
Asteraceae Family ◽  
Positive Results ◽  
Commercial Kit

Simple, fast, low-cost and high throughput protocols are required for DNA isolation of plant species. In this study, phenol chloroform isoamyl alcohol and commercial (Sigma) DNA isolation kit methods were applied on some Inula species that belong to Asteraceae family. Genomic DNA amounts, A260, A280, A260/A230 and purity degrees (A260/A280) that were obtained through both methods were measured through electrophoresis and spectrophotometer. Additionally, PCR amplification was realized by primer pairs specific to nrDNA ITS, cpDNA ndhF (972F-1603R) and trnL-F regions. Results showed that maximum genomic DNA in nanograms obtained by phenol chloroform isoamyl alcohol method. The study also revealed that I. macrocephala had the maximum DNA and I. heterolepis had the minimum DNA amount. A260/A280 purity degrees showed that the highest and lowest purity in gDNAs obtained through phenol-choloform isoamyl alcohol method were in I.aucheriana and I. salicina, respectively. The highest and lowest purity degrees of gDNAs obtained through commercial kit was observed in I. fragilis and I. macrocephala samples, respectively. PCR amplification results showed that while band profiles of each three regions (ITS, trnL-F and ndhF) did not yield positive results in PCR amplifications using phenol-choloform isoamyl alcohol method; PCR band profiles obtained through commercial kit yielded positive results. As a result, it is fair to say that the relation of genomic DNA with PCR was found to be more efficient although the maximum amount of genomic DNA was obtained through phenol chloroform isoamyl alcohol method. 

Rapid and High Quality DNA Isolation from Origanum onites for RAPD and ISSR Analysis

2008 ◽  
Vol 63 (7-8) ◽  
pp. 595-598 ◽  
Author(s):  
Emel Sözen ◽  
Ismail Poyraz
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Leaf Tissue ◽  
Isoamyl Alcohol ◽  
Molecular Study ◽  
Inter Simple Sequence Repeat ◽  
Limiting Factor ◽  
Hexadecyltrimethylammonium Bromide ◽  
Genomic Dna Isolation ◽  
Origanum Onites

Origanum onites is an economically important medicinal plant with high essential oil content. Lack of an appropriate DNA isolation procedure is a limiting factor for any molecular study of this plant. We have used a protocol for genomic DNA isolation based on a hexadecyltrimethylammonium bromide (CTAB) method described for other plant species. The method involves mortar grinding of leaf tissue, modified CTAB extraction using high salt concentrations and polyvinyl pyrrolidone, and successive isoamyl alcohol/chloroform extractions. The yield was approx. 20 μg DNA per 200 mg of initial fresh plant material. The genomic DNA obtained by this method was suitable to be used in restriction digests, inter simple sequence repeat (ISSR) and randomly amplified polymorphic DNA (RAPD) reactions. This extraction method should facilitate the molecular analysis of Origanum chemotypes.

scholarly journals Genomic DNA Isolation and Purification of Two Endemic Medicinal Plants (Pterocarpus Santalinus Linn.F & Pimpinella Tirupatiensis Bal&Subr) of Seshachalam Hills, Tirumala.

2012 ◽  
Vol 4 (1) ◽  
Author(s):  
S. Vipranarayana ◽  
T.N.V.K. V.Prasad ◽  
A. Rajinikanth ◽  
T. Damodharam
Keyword(s):  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Low Cost ◽  
Isolation And Purification ◽  
Genomic Dna Isolation ◽  
Pterocarpus Santalinus ◽  
Ctab Method ◽  
Endemic Medicinal Plant ◽  
High Level

Ptercarpus santalinus is an important endemic medicinal plant with high medicinal values that interfere with DNA extraction procedures and qualitative, quantitative agaros gel electrophoresis. An effective and low-cost protocol for isolating genomic DNA from the roots of Pterocarpus santilinus and Pimpinella tirupatiensis was described in this paper. It involved a modified CTAB method with distilled water pretreating samples.  The A260/A280 absorbance ratio of extracted DNA was found to be free from polysaccharide, poly-phenols and tannins contaminants ranged from 2.2 to 2.8 within the high level of purity.

A Low-Cost High-Throughput Method for Plant Genomic DNA Isolation

2019 ◽  
pp. 1-7
Author(s):  
Prateek Gupta ◽  
Hymavathi Salava ◽  
Yellamaraju Sreelakshmi ◽  
Rameshwar Sharma
Keyword(s):  
High Throughput ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Low Cost ◽  
Genomic Dna Isolation ◽  
High Throughput Method

scholarly journals Optimizing the pure genomic DNA isolation procedure for Plectranthus amboinicus DNA – A prerequisite for further genomic Studies

2017 ◽  
Vol 2 (4) ◽  
Author(s):  
S. Manikandan ◽  
S. Ansarali ◽  
G.M. Alagu Lakshmanan
Keyword(s):  
Dna Barcoding ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Leaf Tissue ◽  
Isolation Procedure ◽  
Modified Method ◽  
Genomic Dna Isolation ◽  
Young Leaves ◽  
Plectranthus Amboinicus

The DNA isolation procedure for different plant groups have been studied and standardized.  The isolation of pure genomic DNA is the most essential component for any type of molecular studies.  The present work is aimed to identify suitable DNA markers for the amplification of P.amboinicus  DNA  becomes a great hurdle  for DNA barcoding studies carried out by rbcL and matK primers Used in the members of Lamiaceae. To solve this problem, The DNA was extracted by three methods from fresh young leaf tissue of P.amboinicus. After the evaluationthe outcome of these methods, one most suitable modified method was selected for isolating DNA from young leaves of P.amboinicus and selected for suitable DNA barcoding markers for PCR amplification. The quality and quantity of DNAs are a prerequisite for genetic studies for a variety of plants including P.amboinicus. The quantity and quality of the DNA extracted by this method wasused for suitable DNA barcoding markers selection.

scholarly journals Optimizing Genomic DNA Isolation and PCR Amplification For Pasak Bumi (Eurycoma longifolia)

2019 ◽  
Vol 1 (2) ◽  
pp. 30
Author(s):  
Arida Susilowati ◽  
Henti Hendalastuti Rachmat ◽  
Ahmad Baiquni Rangkuti ◽  
Deni Elfiati ◽  
Ami Ambarwati
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Eurycoma Longifolia ◽  
Genomic Dna Isolation

.

scholarly journals Development of a Competent and Trouble Free DNA Isolation Protocol for Downstream Genetic Analyses in Glycine Species

2016 ◽  
Vol 4 (8) ◽  
pp. 700 ◽  
Author(s):  
Muhammad Amjad Nawaz ◽  
Faheem Shehzad Baloch ◽  
Hafiz Mamoon Rehman ◽  
Bao Le ◽  
Fahima Akther ◽  
...  
Keyword(s):  
Molecular Biology ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Plant Molecular Biology ◽  
Pcr Amplification ◽  
Cost Effective ◽  
Extraction Methods ◽  
Glycine Species ◽  
Genomic Dna Isolation

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.

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

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