scholarly journals A Simple Method for DNA Extraction from Mature Date Palm Leaves: Impact of Sand Grinding and Composition of Lysis Buffer

2010 ◽  
Vol 11 (9) ◽  
pp. 3149-3157 ◽  
Author(s):  
Ibrahim A. Arif ◽  
Mohammad A. Bakir ◽  
Haseeb A. Khan ◽  
Anis Ahamed ◽  
Ahmad H. Al Farhan ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Date Palm ◽  
Simple Method ◽  
Lysis Buffer

scholarly journals A Rapid Bacteriophage DNA Extraction Method

2018 ◽  
Vol 1 (3) ◽  
pp. 27 ◽  
Author(s):  
Džiuginta Jakočiūnė ◽  
Arshnee Moodley
Keyword(s):  
Dna Extraction ◽  
Illumina Miseq ◽  
Extraction Methods ◽  
Resistant Bacteria ◽  
Simple Method ◽  
Antibiotic Resistant ◽  
Phage Dna ◽  
Specialized Equipment ◽  
Miseq Platform ◽  
Dna Extraction Methods

Bacteriophages (phages) are intensely investigated as non-antibiotic alternatives to circumvent antibiotic resistance development as well as last resort therapeutic options against antibiotic resistant bacteria. As part of gaining a better understanding of phages and to determine if phages harbor putative virulence factors, whole genome sequencing is used, for which good quality phage DNA is needed. Traditional phage DNA extraction methods are tedious and time consuming, requiring specialized equipment e.g., an ultra-centrifuge. Here, we describe a quick and simple method (under four hours) to extract DNA from double stranded DNA (dsDNA) phages at titers above 1.0 × 1010 plaque-forming units (PFU)/mL. This DNA was suitable for library preparation using the Nextera XT kit and sequencing on the Illumina MiSeq platform.

scholarly journals A modified SDS-based DNA extraction method from raw soybean

2019 ◽  
Vol 39 (2) ◽  
Author(s):  
Yimiao Xia ◽  
Fusheng Chen ◽  
Yan Du ◽  
Chen Liu ◽  
Guanhao Bu ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Monitoring Methods ◽  
Detection Techniques ◽  
Dna Yield ◽  
Seed Flour ◽  
Dna Extraction Method ◽  
Lysis Buffer

Abstract Soybean is the most important genetically modified (GM) oilseed worldwide. Regulations relating to the approval of biotech soybean varieties and product labeling demand accurate and reliable detection techniques to screen for GM soya. High-quality extracted DNA is essential for DNA-based monitoring methods. Thus, four widely used protocols (SDS, CTAB, DP305, and DNeasy Plant Mini Kit) were compared in the present study to explore the most efficient DNA extraction method for raw soya matrix. The SDS-based method showed the highest applicability. Then crucial factors influencing DNA yield and purity, such as SDS lysis buffer component concentrations and organic compounds used to isolate DNA, were further investigated to improve the DNA obtained from raw soybean seeds, which accounts for the innovation of this work. As a result, lysis buffer (2% SDS (w/v), 150 mM NaCl, 50 mM Tris/HCl, 50 mM EDTA, pH 8.0) and organic reagents including chloroform/isoamyl alcohol (24:1, v/v) (C: I), isopropanol, and ethanol corresponding to the extraction and first and second precipitation procedures, respectively, were used in the optimized SDS method. The optimized method was verified by extracting approximately 2020–2444 ng DNA/mg soybean with A260/280 ratios of 1.862–1.954 from five biotech and non-biotech soybean varieties. Only 0.5 mg of soya was required to obtain enough DNA for PCR amplification using the optimized SDS-based method. These results indicate that the screening protocol in the present study achieves the highest suitability and efficiency for DNA isolation from raw soya seed flour.

scholarly journals Comparison of DNA extraction methods for Aspergillus fumigatus using real-time PCR

2006 ◽  
Vol 55 (9) ◽  
pp. 1187-1191 ◽  
Author(s):  
Lisa J. Griffiths ◽  
Martin Anyim ◽  
Sarah R. Doffman ◽  
Mark Wilks ◽  
Michael R. Millar ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Diagnostic Technique ◽  
Extraction Methods ◽  
Simple Method ◽  
Bead Beating ◽  
Fungal Dna ◽  
Dna Extraction Methods ◽  
High Level

Newer methods such as PCR are being investigated in order to improve the diagnosis of invasive aspergillosis. One of the major obstacles to using PCR to diagnose aspergillosis is a reliable, simple method for extraction of the fungal DNA. The presence of a complex, sturdy cell wall that is resistant to lysis impairs extraction of the DNA by conventional methods employed for bacteria. Numerous fungal DNA extraction protocols have been described in the literature. However, these methods are time-consuming, require a high level of skill and may not be suitable for use as a routine diagnostic technique. Here, a number of extraction methods were compared: a freeze–thaw method, a freeze–boil method, enzyme extraction and a bead-beating method using Mini-BeadBeater-8. The quality and quantity of the DNA extracted was compared using real-time PCR. It was found that the use of a bead-beating method followed by extraction with AL buffer (Qiagen) was the most successful extraction technique, giving the greatest yield of DNA, and was also the least time-consuming method assessed.

scholarly journals Determination of production capacity of circulated primordial germ cells (circulated-PGCs) of KUB chicken using lysis buffer ammonium chloride potassium (ACK)

2016 ◽  
Vol 21 (1) ◽  
pp. 55
Author(s):  
Soni Sopiyana ◽  
Iman Supriatna ◽  
M. Agus Setiadi ◽  
Mohamad Fahrudin
Keyword(s):  
Embryonic Development ◽  
Ammonium Chloride ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Production Capacity ◽  
Development Stage ◽  
Simple Method ◽  
Average Production ◽  
Short Period ◽  
Lysis Buffer

<p class="abstrak2">In poultry embryos, primordial germ cells (PGCs) are progenitor cells for gametes, which have unique migration pathway. Primordial germ cells arise from epiblast in germinal crescent and circulate through the bloodstream for a short period of time, then leave blood vessel to migrate toward gonads. The aim of this study was to determine the potential production capacity of circulated-PGCs of KUB chicken at different developmental stages of embryo using a rapid and simple method. Seventy five KUB chicken fertile eggs were divided into five groups and incubated at 38.5 <sup>0</sup>C with a humidity of 60%. Hatching was set to the embryonic development stage of 14-18. The blood was collected through dorsal aorta using micropipette under microscope. The collected blood was placed in a 1.5 ml eppendorf tube which was previously filled with 100 µl phosphate buffered saline without Ca<sup>2+</sup> and Mg<sup>2+</sup> (PBS-) mixed with fetal bovine serum (FBS) with a ratio of 90%:10%. The PGCs were purified using lysis buffer ammonium chloride potassium method. The results showed that average production of circulated-PGCs per embryo of KUB chicken were significantly affected by stage of embryonic development (P &lt;0.05). The average production of circulated-PGCs at stage 14, 15, 16, 17, and 18 were 37.9; 53.5; 49.8; 38.3; and 33.5 respectively. The number of circulated-PGCs was not different among stages 14, 17 nor 18. The highest number of circulated-PGCs of KUB chicken was obtained at stage 15, so that the isolation and collection of PGCs through the blood circulation was recommended in stage 15.</p><strong>Key Words: </strong>KUB Chicken, PGCs, Embryonic Development Stage, Ammonium Chloride Potassium

A Simple Method of DNA Extraction and STR Typing from Urine Samples Using a Commercially Available DNA/RNA Extraction Kit

2003 ◽  
Vol 48 (1) ◽  
pp. 2002184 ◽  
Author(s):  
Toshihiro Yasuda ◽  
Reiko Iida ◽  
Haruo Takeshita ◽  
Misuzu Ueki ◽  
Tamiko Nakajima ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Rna Extraction ◽  
Urine Samples ◽  
Str Typing ◽  
Simple Method

A rapid and simple method for small-scale DNA extraction inAgavaceae and other tropical plants

2002 ◽  
Vol 20 (3) ◽  
pp. 299-299 ◽  
Author(s):  
Miguel Keb-Llanes ◽  
Gerardo González ◽  
Bartolomé Chi-Manzanero ◽  
Diógenes Infante
Keyword(s):  
Dna Extraction ◽  
Small Scale ◽  
Simple Method ◽  
Tropical Plants
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