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 Extraction of high-quality genomic DNA and identification of different DNA barcoding markers for chickpea (Cicer arietinum L.)

2020 ◽  
Vol 1 (1) ◽  
pp. 7
Author(s):  
Umavathi Saraswathi ◽  
Lakshmanan Mullainathan
Keyword(s):  
Dna Barcoding ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Genomic Library ◽  
Pcr Amplification ◽  
Slight Modification ◽  
High Quality ◽  
Universal Primer ◽  
Genetic Studies

The genetic studies of individual plants, especially self-pollinated species like chickpea need to be evaluated at the DNA level with the help of molecular markers for identifying genetic variations among the plants. High-quality DNA extraction is a prerequisite for genetic studies. Extraction of intact genomic DNA with high – molecular mass is essential for the study of many molecular biology applications like Polymerase Chain Reaction, endonuclease restriction digestion, southern blot analysis, and also for the construction of a genomic library. Several plant DNA extraction methods are available, even though the DNA isolation methods that give good yield employing both quantity and quality is quite difficult especially for self-pollinated crops like a chickpea. This work was focused on developing a standard protocol for the extraction of genomic DNA and identifying different barcoding markers. The result revealed that the CTAB extraction method with slight modification in protocol had been optimized for DNA isolation. The purified DNA, which was isolated through the CTAB method, had excellent spectral qualities and is efficiently digested by a restriction endonuclease, and is found to be more suitable for long-fragment PCR amplification. DNA barcoding is considered as a promising tool because it provides a practical and standard identification of plants. The isolated DNA sample was processed with a classical DNA barcoding approach by amplifying and sequencing with a universal primer. According to the result, among the different barcoding markers studied, the RbcL and Mat K were found to given the best result for molecular species identification in chickpea.

scholarly journals Optimalisasi Teknik Ekstraksi dan Isolasi DNA Tanaman Suren (Toona Sureni Merr.) untuk Analisis Keragaman Genetik berdasarkan Random Amplified Polymorphic DNA (RAPD)

2012 ◽  
Vol 14 (1) ◽  
pp. 138 ◽  
Author(s):  
Muh Restu ◽  
Mukrimin Mukrimin ◽  
Gusmiaty Gusmiaty
Keyword(s):  
Genomic Dna ◽  
Extraction Method ◽  
Dna Isolation ◽  
Random Amplified Polymorphic Dna ◽  
Pcr Amplification ◽  
Secondary Compounds ◽  
Extraction Methods ◽  
Extraction Techniques ◽  
High Quality ◽  
Optimum Extraction

The species of trees have different secondary compounds that need optimum extraction techniques. Appropriate extraction techniquesdetermine the quality and quantity of DNA produced. This research aims to found optimal of extraction methods and DNA isolation, thento created genome DNA in high quality and quantity, so that it can be using for genetic variation analyses in Suren (Toona sureni Merr.) byRandom Amplified Polymorphic DNA (RAPD). This study shows that DNA concentrates were 763.3 μg/ml, 180.0 μg/ml, 383.3 μg/ml, and436.7 μg/ml. While based on the results of PCR amplification using the primers OPD 03 shows that the four extraction methods used, the extraction method of number 3 has been able to produce genomic DNA with better quality and more number of bands, although the quantityis lower.

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 A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis

2020 ◽  
Author(s):  
Wei Hu ◽  
J. Clark Lagarias
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Method ◽  
Low Cost ◽  
Plant Molecular Biology ◽  
Pcr Analysis ◽  
High Quality ◽  
Plant Dna ◽  
Genomic Dna Isolation ◽  
Dna Extraction Method

AbstractBackgroundConsistent isolation of high quality plant genomic DNA is a prerequisite for successful PCR analysis. Time consumption, ease of operation and procedure cost are important secondary considerations for selecting an effective DNA extraction method. The simple, reliable and rapid DNA extraction method developed by Edwards and colleagues in 1991 [1] has proven to be the gold standard.ResultsThrough modification of the Edwards method of extraction, we have developed a one-tube protocol that greatly improves the efficiency of plant DNA extraction and reduces the potential for sample contamination while simultaneously yielding high quality DNA suitable for PCR analysis. We further show that DNA extracts prepared with this method are stable at room temperature for at least three months.ConclusionThe one-tube extraction method yields high quality plant DNA with improved efficiency while greatly minimizing the potential for cross contamination. This low-cost and environment-friendly method is widely applicable for plant molecular biology research.

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 Novel kit for efficient extraction of long-segment genomic DNA by nanodisk adsorption

2019 ◽  
Vol 1 (1) ◽  
pp. 4-5
Author(s):  
Siqi Wu ◽  
Zhaofang Han ◽  
Hesong Qiu
Keyword(s):  
Molecular Biology ◽  
Dna Extraction ◽  
High Purity ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Biochemical Properties ◽  
High Efficacy ◽  
Tissue Samples ◽  
Efficient Extraction ◽  
Total Dna

The acquisition of complete DNA is usually the first and most critical step in many basic molecular biology applications. Isolation of complete total DNA from tissue samples is related to different physical and biochemical properties of tissue. Long fragments of DNA are easily degraded and fragmented, which poses a challenge for extracting complete long fragments of DNA. By comparing several different extraction methods and verifying the concentration and length of extracted DNA, it is proved that the kit we used has high  efficacy. We provide tissue kit DNA extraction for long fragments and obtain long fragments of high purity DNA from almost any type of tissue, especially muscle and blood tissues.

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. 

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