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 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 A rapid and simple bead-bashing-based method for genomic DNA extraction from mammalian tissue

BioTechniques ◽  
2020 ◽  
Vol 68 (5) ◽  
pp. 240-244
Author(s):  
Shan Wei ◽  
Brynn Levy ◽  
Nataly Hoffman ◽  
Claudia Cujar ◽  
Reunet Rodney-Sandy ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Rapid Method ◽  
Human Tissue ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Clinical Applications ◽  
Mammalian Tissue ◽  
Fume Hood ◽  
Molecular Assays ◽  
Tissue Specimens

Conventional genomic DNA (gDNA) extraction methods can take hours to complete, may require fume hoods and represent the most time-consuming step in many gDNA-based molecular assays. We systematically optimized a bead bashing-based (BBB) approach for rapid gDNA extraction without the need for a fume hood. Human tissue specimens (n = 34) subjected to the 12-min BBB method yielded 0.40 ± 0.17 (mean ± SD) μg of gDNA per milligram of tissue, sufficient for many downstream applications, and 3- and 6-min extensions resulted in an additional 0.43 ± 0.23 μg and 0.48 ± 0.43 μg per milligram of tissue, respectively. The BBB method provides a simple and rapid method for gDNA extraction from mammalian tissue that is applicable to time-sensitive clinical applications.

scholarly journals Evaluation of the impact of six different DNA extraction methods for the representation of the microbial community associated with human chronic wound infections using a gel-based DNA profiling method

AMB Express ◽  
2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Ayomi Dilhari ◽  
Asanga Sampath ◽  
Chinthika Gunasekara ◽  
Neluka Fernando ◽  
Deepaka Weerasekara ◽  
...  
Keyword(s):  
Microbial Community ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Chronic Wounds ◽  
Chronic Wound ◽  
Extraction Methods ◽  
Wound Infections ◽  
Dna Extraction Methods ◽  
The Impact ◽  
Chronic Wound Infections

AbstractInfected chronic wounds are polymicrobial in nature which include a diverse group of aerobic and anaerobic microorganisms. Majority of these communal microorganisms are difficult to grow in vitro. DNA fingerprinting methods such as polymerase chain reaction-denaturation gradient gel electrophoresis (PCR-DGGE) facilitate the microbial profiling of complex ecosystems including infected chronic wounds. Six different DNA extraction methods were compared for profiling of the microbial community associated with chronic wound infections using PCR-DGGE. Tissue debris obtained from chronic wound ulcers of ten patients were used for DNA extraction. Total nucleic acid was extracted from each specimen using six DNA extraction methods. The yield, purity and quality of DNA was measured and used for PCR amplification targeting V2–V3 region of eubacterial 16S rRNA gene. QIAGEN DNeasy Blood and Tissue Kit (K method) produced good quality genomic DNA compared to the other five DNA extraction methods and gave a broad diversity of bacterial communities in chronic wounds. Among the five conventional methods, bead beater/phenol–chloroform based DNA extraction method with STES buffer (BP1 method) gave a yield of DNA with a high purity and resulted in a higher DGGE band diversity. Although DNA extraction using heat and NaOH had the lowest purity, DGGE revealed a higher bacterial diversity. The findings suggest that the quality and the yield of genomic DNA are influenced by the DNA extraction protocol, thus a method should be carefully selected in profiling a complex microbial community.

Genomic DNA extraction methods using formalin-fixed paraffin-embedded tissue

2015 ◽  
Vol 486 ◽  
pp. 17-23 ◽  
Author(s):  
Keerti Potluri ◽  
Ahmed Mahas ◽  
Michael N. Kent ◽  
Sameep Naik ◽  
Michael Markey
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Genomic Dna Extraction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Dna Extraction Methods ◽  
Formalin Fixed

scholarly journals Comparison of DNA Extraction Methods for Metagenomic Sequencing in Anaerobic Digestion

2021 ◽  
Vol 43 (1) ◽  
pp. 43-50
Author(s):  
Juhee Shin ◽  
Youngback Kim ◽  
Seung Gu Shin
Keyword(s):  
Microbial Community ◽  
South Korea ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Microbial Populations ◽  
Metagenomic Sequencing ◽  
Unifrac Distance ◽  
Commercial Kits ◽  
Dna Extraction Methods

Objectives:This study was performed to compare three commercial kits for the extraction of genomic DNA from anaerobic digestate for subsequent iSeq 100 sequencing and microbial community analysis.Methods:A full-scale mesophilic biogas plant was sampled, and made into aliquots of identical volumes to extract DNA using three commercial kits: FastDNA spin kit for soil (MP Biomedicals, USA), Exgene stool DNA mini (GeneAll, South Korea) and AccuPrep genomic DNA extraction kit (Bioneer, South Korea). To analyze the microbial communities in the purified DNA, 16S rDNA amplicon sequencing (V3-4 region for bacteria and V4-5 region for archaea) was performed on the Illumina iSeq 100 platform. Quality filtered sequence reads were OTU-clustered for taxonomic assignment conducted using the RDP classifier on-line.Results and Discussion:The microbial community structure visualized on the NMDS plot using the weighted UniFrac distance revealed that both bacteria and archaeal communities have phylogenetic differences depending on the DNA extraction kit used. In addition, the abundance of certain microbial populations was significantly different among the DNA extraction methods. For example, Proteobacteria was the least abundant using the soil kit, while this phylum was the most abundant when the stool kit was used. However, in the case of Thermotogae, this tendency was vice versa. The abundance of archaeal genera Methanomethylovorans and Methanosarcina was also affected by DNA extraction methods.Conclusions:The microbial populations analyzed by 16S based sequencing were affected by DNA extraction methods. To compare microbial community changes in the identical set of research, one DNA extraction method should be chosen and used consistently for the whole experiment.

scholarly journals Comparison of three genomic DNA extraction methods from soursop leaves (Annona muricata L.)

2020 ◽  
Vol 5 (2) ◽  
pp. 106-119
Author(s):  
Rosalba Lira-Ortiz ◽  
◽  
Moisés Alberto Cortés-Cruz ◽  
Blanca Amalia Amaro- González ◽  
Graciela Guadalupe López-Guzmán ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Annona Muricata ◽  
Genomic Dna Extraction ◽  
Dna Extraction Methods
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