Leaf Tissue Sampling and DNA Extraction Protocols

2013 ◽  
pp. 53-67 ◽  
Author(s):  
Kassa Semagn
Keyword(s):  
Dna Extraction ◽  
Leaf Tissue ◽  
Tissue Sampling

scholarly journals Comparison of genomic DNA extraction methods to obtain high DNA quality from barnyard grass (Echinochloa colona)

2020 ◽  
Author(s):  
Romesh Kumar Salgotra ◽  
Bhagirath Singh Chauhan
Keyword(s):  
Essential Oils ◽  
Dna Extraction ◽  
Leaf Tissue ◽  
Extraction Methods ◽  
High Quality ◽  
Time Saving ◽  
Plant Leaf ◽  
Plant Parts ◽  
Molecular Studies ◽  
Leaf Tissues

Abstract Background: The study of weed genomics is important for the effective management of weeds to enhance crop yield. A rapid, inexpensive and high quality DNA extraction is needed for genomic and other molecular studies. Here, we describe the protocols for DNA extraction from two different parts of the Echinochloa colona plant using modified cetyltrimethylammonium bromide (CTAB) and a commercial kit.Results: In the study, it was observed that the DNA extracted from plant leaf tissues and dry seeds with a modified CTAB protocol was of good quality, with no contaminations of polysaccharides and essential oils. Quality of DNA extracted from dry seeds was comparable with that of plant leaves under both protocols. The extracted DNA from both plant parts was successfully amplified by PCR using the EPSPS microsatellite marker. Compared to the protocol of DNA extracted from leaf tissue, dry seeds will save time and other valuable resources. Moreover, the same protocols can be implemented for the extraction of high-quality DNA for molecular studies in other plant species where a large amount of polysaccharides, secondary metabolites and essential oils are present.Conclusions: Modified methods of DNA extraction from dry seeds are efficient and time-saving which can be used in genotypic and other molecular approaches. High-quality DNA can be isolated from plant leaf tissues using modified CTAB and commercial kits, however, DNA extracted from dry seeds will save time and other valuable resources.

scholarly journals Sampling Tissue for DNA Analysis of Trees: Trunk Cambium as an Alternative to Canopy Leaves

2005 ◽  
Vol 54 (1-6) ◽  
pp. 265-269 ◽  
Author(s):  
N. Colpaert ◽  
S. Cavers ◽  
E. Bandou ◽  
H. Caron ◽  
G. Gheysen ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Tree Species ◽  
Dna Analysis ◽  
Leaf Tissue ◽  
Ground Level ◽  
Tropical Tree ◽  
Aflp Analysis ◽  
Simple Method ◽  
Tropical Tree Species ◽  
High Concentrations

Abstract The number of studies of tropical tree species that use molecular tools is increasing, most of which collect leaf tissue for genomic DNA extraction. In tropical trees the canopy is not only frequently inaccessible, but also, once reached, the leaf tissue is often heavily defended against herbivory by high concentrations of anti-predation compounds, which may inhibit downstream applications, particularly PCR. Cambium tissue, accessed directly from the tree trunk at ground level, offers a readily accessible resource that is less hampered by the presence of defensive chemicals than leaf tissue. Here we describe a simple method for obtaining tissue from the cambial zone for DNA extraction and test the applicability of the method in a range of tropical tree species. The method was used successfully to extract DNA from 11 species in nine families. A subset of the DNA extracts was tested in more detail and proved to be highly suitable for AFLP analysis.

An improved method to extract DNA from mango Mangifera indica

Biologia ◽  
2014 ◽  
Vol 69 (2) ◽  
Author(s):  
Mohammad Uddin ◽  
Wenli Sun ◽  
Xinhua He ◽  
Jaime Teixeira da Silva ◽  
Qi Cheng
Keyword(s):  
Secondary Metabolites ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Mangifera Indica ◽  
Leaf Tissue ◽  
Poor Quality ◽  
Ammonium Bromide ◽  
High Quality ◽  
Woody Perennials ◽  
Ctab Method

AbstractHigh quality genomic DNA is the first step in the development of DNA-based markers for fingerprinting and genetic diversity of crops, including mango (Mangifera indica L.), a woody perennial. Poor quality genomic DNA hinders the successful application of analytical DNA-based tools. Standard protocols for DNA extraction are not suitable for mango since the extracted genomic DNA often contains secondary metabolites that interfere with analytical applications. In this study, we employed an additional step to remove polysaccharides, polyphenols and secondary metabolites from genomic DNA extracted from young or mature leaf tissue; then a modified traditional cetyl trimethyl ammonium bromide (CTAB) method was applied. The use of 0.4 M glucose improved DNA quality and avoided contamination and browning by polyphenolics, relative to the traditional CTAB method. This is an easy and efficient method for genomic DNA extraction from both young and mature leaves of mango. The isolated DNA was free of polysaccharides, polyphenols, RNA and other major contaminants, as judged by its clear colour, its viscosity, A260/A280 ratio and suitability for PCR-based reactions. This modified protocol was also used to extract high quality genomic DNA from other woody perennials, including walnut, guava, lychee, pear, grape and sugarcane.

scholarly journals Identifying Buffalograss [Buchloe dactyloides (Nutt.) Engelm.] Cultivar Breeding Lines Using Random Amplified Polymorphic DNA (RAPD) Markers

1994 ◽  
Vol 119 (1) ◽  
pp. 126-130 ◽  
Author(s):  
Lin Wu ◽  
Hong Lin
Keyword(s):  
Dna Extraction ◽  
Rapd Markers ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Sodium Dodecyl ◽  
Leaf Tissue ◽  
Extraction Methods ◽  
Breeding Lines ◽  
Random Primers ◽  
Dna Extraction Methods

The polymerase chain reaction (PCR) and RAPD fragments are potentially useful methods for identifying turfgrass cultivar breeding lines. RAPD markers were studied in 25 vegetatively propagated buffalograss lines using oligonucleotide random primers and agarose-gel electrophoresis to determine their potential for identifying cultivar breeding lines. The variation of RAPD markers was extensive. The RAPD markers produced by one random primer were sufficient to separate the 25 buffalograss lines. Cluster analysis baaed on' the RAPD markers produced by two random primers revealed that the 25 buffalograss lines generally fell into two groups: diploid and hexaploid. Three DNA extraction methods—sarcosyl lysis-chloroform extraction-isopropanol precipitation, sodium dodecyl sulfate (SDS) lysine-isopropanol precipitation, and boiling in the presence of Chelex-100 resin—and fresh or oven-dried tissues were tested for reproducibility of RAPD markers. The three DNA extraction methods, using dry or fresh plant tissues, produced highly comparable RAPD marker profiles. More than 80%1 of the RAPD markers was consistently detected in six replicate analyses. The above studies demonstrate that small quantities (5 mg) of oven-dried leaf tissue and several DNA extraction methods can be used for buffalograss fingerprint studies.

scholarly journals Reliability of non-invasive tissue sampling methods for DNA extraction in rabbits (Oryctolagus cuniculus)

2012 ◽  
Vol 20 (2) ◽  
Author(s):  
Manel BEN LARBI ◽  
A. Tircazes ◽  
K. Feve ◽  
F. Tudela ◽  
G. Bolet
Keyword(s):  
Dna Extraction ◽  
Oryctolagus Cuniculus ◽  
Sampling Methods ◽  
Tissue Sampling ◽  
Non Invasive

scholarly journals Sugarcane Leaf Tissue Sample Preparation for Diagnostic Analysis

EDIS ◽  
2019 ◽  
Vol 2005 (15) ◽  
Author(s):  
Ike V. Ezenwa ◽  
J. Mabry McCray ◽  
Peter R. Newman ◽  
Ronald W. Rice
Keyword(s):  
Sample Preparation ◽  
Tissue Sample ◽  
Leaf Tissue ◽  
Publication Date ◽  
Diagnostic Analysis ◽  
Tissue Sampling ◽  
Original Publication ◽  
Nutrient Analysis ◽  
Foliar Nutrient

This document focuses on sugarcane leaf tissue sampling for the purpose of foliar nutrient analysis and interpretation. This document is SS-AGR-259, one of a series of the Agronomy Department, UF/IFAS Extension. Original publication date November 2005. SS-AGR-259/SC076: Sugarcane Leaf Tissue Sample Preparation for Diagnostic Analysis (ufl.edu)

scholarly journals Comparison of Different Leaf Preservation Methods To Obtain High Quality DNA From Enset (Ensete Ventricosum (Welw.) Cheesman), A Native And Orphan Food Security Crop In Ethiopia

2021 ◽  
Author(s):  
Alye Tefera Haile ◽  
Sylvia Sagen Johnsen ◽  
Mallikarjuna Rao Kovi ◽  
Trine Hvoslef-Eide ◽  
Bizuayehu Tesfaye ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Silica Gel ◽  
Genomic Dna ◽  
Leaf Tissue ◽  
Endemic Plant ◽  
High Quality ◽  
Crop Species ◽  
Ensete Ventricosum ◽  
Ctab Solution ◽  
Ctab Method

Abstract Background: Enset (Ensete ventricosum (Welw.) Cheesman) is a staple food for more than 20 million Ethiopians and only cultivated in the native indigenous farming systems of Ethiopia. In contrast to other cultivated species in the Musaceae family, enset has been relatively little studied at the molecular level. Application of advanced molecular genetic techniques requires rapid extraction of DNA of high quality and quantity. Fresh, lyophilized tissues, as well as tissues stored in liquid nitrogen are mainly preferred to avoid DNA degradation, thus most of the DNA extraction protocols recommend these types of tissues as starting material. However, such sample processing techniques are difficult to utilize in many developing countries and at collection sites of many endemic plant species, underutilized or orphan crop species like enset. These situations necessitate the development of alternative protocols for leaf preservation and optimized methods for isolating high-quality DNA from dried or preserved leaf samples. Results: In this study, three different leaf preservation and two DNA extraction methods were compared. Fresh young leaf tissue was preserved using the minor modified saturated NaCl-CTAB solution, silica gel or 96% ethanol at ambient temperature for more than 35 days. Subsequently, DNA was extracted using either the DNeasy Plant Mini Kit or the CTAB method. As compared to silica gel and 96% ethanol, the minor modified saturated NaCl-CTAB solution preserved the quality, quantity, and integrity of enset genomic DNA. This method consistently produced genomic DNA of high-quality and quantity at affordable cost. The DNeasy Plant Mini Kit method was found to be more efficient than the standard CTAB method, being faster and producing genomic DNA of higher quality. Conclusions: Using saturated NaCl-CTAB solution is an accessible, efficient, scalable, and inexpensive way to preserve enset leaves during collection and transportation. The preservation protocol was validated for leaf tissues of all cultivated and wild enset, and Entada landraces. Genomic DNA of high quality and quantity was obtained from preserved enset leaves, which can be used for further downstream applications including PCR and sequencing.

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