scholarly journals Research Article A modified non-liquid nitrogen protocol for extraction of high-quality genomic DNA from the inner bark tissues of Dalbergia cochinchinensis (Fabaceae)

2021 ◽  
Vol 20 (2) ◽  
Author(s):  
X.C. Mo ◽  
P.P. Wangsomnuk
Keyword(s):  
Liquid Nitrogen ◽  
Genomic Dna ◽  
High Quality ◽  
Inner Bark ◽  
Research Article

Extraction of Genomic DNA from Inner Bark of Cedrus deodara (Roxb.) G. Don for RAPD Study

2017 ◽  
Vol 40 (1) ◽  
pp. 25-27
Author(s):  
Akhilesh Kumar ◽  
◽  
Santan Barthwal ◽  
H.S. Ginwal ◽  
◽  
...  
Keyword(s):  
Sodium Chloride ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Rapd Markers ◽  
High Concentration ◽  
High Quality ◽  
Cedrus Deodara ◽  
Remote Areas ◽  
Inner Bark ◽  
Random Amplification

Genomic DNA extraction from forestry tree species require young leaf samples to obtain high-quality DNA for molecular based study. For some study, leaf samples must be collected from remote areas and are difficult to transport long distances. We developed alternative method of DNA extraction from inner bark of Cedrus deodara. We used 2.5% PVP for removal of phenolic compound and apply high concentration of sodium chloride to removes polysaccharides. Extracted DNA gives positive amplification with PCR using random amplification of polymorphic DNA (RAPD) markers.

scholarly journals DNA Extraction Protocol for Plants with High Levels of Secondary Metabolites and Polysaccharides without Using Liquid Nitrogen and Phenol

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Sunil Kumar Sahu ◽  
Muthusamy Thangaraj ◽  
Kandasamy Kathiresan
Keyword(s):  
Salt Marsh ◽  
Secondary Metabolites ◽  
Liquid Nitrogen ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Wide Spectrum ◽  
Pcr Amplification ◽  
Restriction Digestion ◽  
High Quality ◽  
Salt Marsh Species

Mangroves and salt marsh species are known to synthesize a wide spectrum of polysaccharides and polyphenols including flavonoids and other secondary metabolites which interfere with the extraction of pure genomic DNA. Although a plethora of plant DNA isolation protocols exist, extracting DNA from mangroves and salt marsh species is a challenging task. This study describes a rapid and reliable cetyl trimethylammonium bromide (CTAB) protocol suited specifically for extracting DNA from plants which are rich in polysaccharides and secondary metabolites, and the protocol also excludes the use of expensive liquid nitrogen and toxic phenols. Purity of extracted DNA was excellent as evident by A260/A280 ratio ranging from 1.78 to 1.84 and A260/A230 ratio was >2, which also suggested that the preparations were sufficiently free of proteins and polyphenolics/polysaccharide compounds. DNA concentration ranged from 8.8 to 9.9 μg μL−1. The extracted DNA was amenable to RAPD, restriction digestion, and PCR amplification of plant barcode genes (matK and rbcl). The optimized method is suitable for both dry and fresh leaves. The success of this method in obtaining high-quality genomic DNA demonstrated the broad applicability of this method.

High Microsatellite and SNP Genotyping Success Rates Established in a Large Number of Genomic DNA Samples Extracted From Mouth Swabs and Genotypes

2006 ◽  
Vol 9 (4) ◽  
pp. 501-506 ◽  
Author(s):  
Josine L. Min ◽  
Nico Lakenberg ◽  
Margreet Bakker-Verweij ◽  
Eka Suchiman ◽  
Dorret I. Boomsma ◽  
...  
Keyword(s):  
Success Rate ◽  
Genomic Dna ◽  
Snp Genotyping ◽  
High Yield ◽  
Success Rates ◽  
Nucleotide Polymorphism ◽  
High Quality ◽  
Single Nucleotide ◽  
Dna Yield ◽  
Yield And Quality

AbstractIn this article, we present the genomic DNA yield and the microsatellite and single nucleotide polymorphism (SNP) genotyping success rates of genomic DNA extracted from a large number of mouth swab samples. In total, the median yield and quality was determined in 714 individuals and the success rates in 378,480 genotypings of 915 individuals. The median yield of genomic DNA per mouth swab was 4.1 μg (range 0.1–42.2 μg) and was not reduced when mouth swabs were stored for at least 21 months prior to extraction. A maximum of 20 mouth swabs is collected per participant. Mouth swab samples showed in, respectively, 89% for 390 microsatellites and 99% for 24 SNPs a genotyping success rate higher than 75%. A very low success rate of genotyping (0%–10%) was obtained for 3.2% of the 915 mouth swab samples using microsatellite markers. Only 0.005% of the mouth swab samples showed a geno-typing success rate lower than 75% (range 58%–71%) using SNPs. Our results show that mouth swabs can be easily collected, stored by our conditions for months prior to DNA extraction and result in high yield and high-quality DNA appropriate for genotyping with high success rate including whole genome searches using microsatellites or SNPs.

scholarly journals A simple ethanol wash of the tissue homogenates recovers high-quality genomic DNA from Corchorus species characterized by highly acidic and proteinaceous mucilages

2001 ◽  
Vol 14 (1) ◽  
Author(s):  
Avijit Kundu ◽  
Debabrata Sarkar ◽  
Amit Bhattacharjee ◽  
Niladri Topdar ◽  
Mohit Kumar Sinha ◽  
...  
Keyword(s):  
Genomic Dna ◽  
High Quality ◽  
Tissue Homogenates

scholarly journals Facile, High Quality Sequencing of Bacterial Genomes from Small Amounts of DNA

2014 ◽  
Vol 2014 ◽  
pp. 1-8
Author(s):  
Momchilo Vuyisich ◽  
Ayesha Arefin ◽  
Karen Davenport ◽  
Shihai Feng ◽  
Cheryl Gleasner ◽  
...  
Keyword(s):  
Genomic Dna ◽  
De Novo ◽  
Gc Content ◽  
Library Preparation ◽  
Sequencing Data ◽  
Bacterial Genomes ◽  
Dna Amount ◽  
High Quality ◽  
Preparation Methods

Sequencing bacterial genomes has traditionally required large amounts of genomic DNA (~1 μg). There have been few studies to determine the effects of the input DNA amount or library preparation method on the quality of sequencing data. Several new commercially available library preparation methods enable shotgun sequencing from as little as 1 ng of input DNA. In this study, we evaluated the NEBNext Ultra library preparation reagents for sequencing bacterial genomes. We have evaluated the utility of NEBNext Ultra for resequencing andde novoassembly of four bacterial genomes and compared its performance with the TruSeq library preparation kit. The NEBNext Ultra reagents enable high quality resequencing andde novoassembly of a variety of bacterial genomes when using 100 ng of input genomic DNA. For the two most challenging genomes (Burkholderiaspp.), which have the highest GC content and are the longest, we also show that the quality of both resequencing andde novoassembly is not decreased when only 10 ng of input genomic DNA is used.

scholarly journals An alternative method for Staphylococcus aureus DNA isolation

2008 ◽  
Vol 60 (2) ◽  
pp. 299-306 ◽  
Author(s):  
L. Chapaval ◽  
D.H. Moon ◽  
J.E. Gomes ◽  
F.R. Duarte ◽  
S.M. Tsai
Keyword(s):  
Staphylococcus Aureus ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Environmental Isolates ◽  
Reaction Volume ◽  
High Quality ◽  
Rapid Procedure ◽  
Final Reaction ◽  
Staphylococcal Enterotoxins ◽  
Final Reaction Volume

This study describes a rapid procedure for the isolation of genomic DNA from Staphylococcus aureus that yielded a good amount of high quality DNA for the amplification of staphylococcal enterotoxins genes (A, B, C, D, and E) and the TSST-1 gene as well as enzymatic restriction (HaeIII) from environmental isolates. With this method, it was possible to detect these genes in a sample containing as little as 10(5) cells with positive PCR reactions obtained from approximately 10pg of DNA in a final reaction volume of 25µl.

scholarly journals Optimization of high molecular weight DNA extraction methods in shrimp for a long-read sequencing platform

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10340
Author(s):  
Pacharaporn Angthong ◽  
Tanaporn Uengwetwanit ◽  
Wirulda Pootakham ◽  
Kanchana Sittikankaew ◽  
Chutima Sonthirod ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Marine Organisms ◽  
High Quality ◽  
Sequencing Platform ◽  
Global Food Security ◽  
Long Read ◽  
Ctab Method ◽  
Global Food

Marine organisms are important to global food security as they are the largest source of animal proteins feeding mankind. Genomics-assisted aquaculture can increase yield while preserving the environment to ensure sufficient and sustainable production for global food security. However, only few high-quality genome sequences of marine organisms, especially shellfish, are available to the public partly because of the difficulty in the sequence assembly due to the complex nature of their genomes. A key step for a successful genome sequencing is the preparation of high-quality high molecular weight (HMW) genomic DNA. This study evaluated the effectiveness of five DNA extraction protocols (CTAB, Genomic-tip, Mollusc DNA, TIANamp Marine Animals DNA, and Sbeadex livestock kits) in obtaining shrimp HMW DNA for a long-read sequencing platform. DNA samples were assessed for quality and quantity using a Qubit fluorometer, NanoDrop spectrophotometer and pulsed-field gel electrophoresis. Among the five extraction methods examined without further optimization, the Genomic-tip kit yielded genomic DNA with the highest quality. However, further modifications of these established protocols might yield even better DNA quality and quantity. To further investigate whether the obtained genomic DNA could be used in a long-read sequencing application, DNA samples from the top three extraction methods (CTAB method, Genomic-tip and Mollusc DNA kits) were used for Pacific Biosciences (PacBio) library construction and sequencing. Genomic DNA obtained from Genomic-tip and Mollusc DNA kits allowed successful library construction, while the DNA obtained from the CTAB method did not. Genomic DNA isolated using the Genomic-tip kit yielded a higher number of long reads (N50 of 14.57 Kb) than those obtained from Mollusc DNA kits (N50 of 9.74 Kb). Thus, this study identified an effective extraction method for high-quality HMW genomic DNA of shrimp that can be applied to other marine organisms for a long-read sequencing platform.

scholarly journals Urine as a high‐quality source of host genomic DNA from wild populations

2020 ◽  
Vol 21 (1) ◽  
pp. 170-182
Author(s):  
Andrew T. Ozga ◽  
Timothy H. Webster ◽  
Ian C. Gilby ◽  
Melissa A. Wilson ◽  
Rebecca S. Nockerts ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Wild Populations ◽  
High Quality
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