Successful and cost-effective DNA extraction method from insects with small amount of tissue v2 (protocols.io.bfajjicn)

Through the advancement of biotechnology, DNA-based methods are the most effective techniques in species identification, as they are rapid and have higher stability in harsh conditions compared to protein-based methods. This study was conducted to determine the efficiency of the traditional DNA extraction method, phenol/chloroform/isoamyl alcohol (PCIA), and comparing it with the commercially available kit by evaluating the purity, concentration, and suitability for amplification of porcine DNA in raw chicken and beef mixtures. The quantity and quality of the DNA extracts were assessed using a UV-Vis spectrophotometer. Polymerase chain reaction (PCR) was performed using species-specific primers targeting mitochondrial DNA cytochrome b (cyt b) gene of chicken (227-bp), beef (274-bp), and pork (398-bp), to confirm the template usability and quality of the DNA extracts. High DNA concentrations and purity were obtained from meat samples extracted using the PCIA method. The visualization of pork DNA on 2% agarose gel was able to detect pork contamination in raw meat mixtures up to minute proportion (1%). The existence of pork in chicken and beef was indicated with the presence of a specific 398-bp DNA band. Thus, the PCIA method can be recommended as a cost-effective and an excellent alternative to more expensive extraction kits in detecting pork DNA in raw meat mixtures.

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Choice of DNA extraction method affects DNA metabarcoding of unsorted invertebrate bulk samples

Metabarcoding and Metagenomics ◽

10.3897/mbmg.2.26664 ◽

2018 ◽

Vol 2 ◽

Cited By ~ 14

Author(s):

Markus Majaneva ◽

Ola H. Diserud ◽

Shannon H.C. Eagle ◽

Mehrdad Hajibabaei ◽

Torbjørn Ekrem

Keyword(s):

Dna Extraction ◽

Extraction Method ◽

Target Material ◽

Cost Effective ◽

Community Analysis ◽

Extraction Methods ◽

Dna Recovery ◽

Dna Yield ◽

Dna Extraction Method ◽

Dna Metabarcoding

Characterisation of freshwater benthic biodiversity using DNA metabarcoding may allow more cost-effective environmental assessments than the current morphological-based assessment methods. DNA metabarcoding methods where sorting or pre-sorting of samples are avoided altogether are especially interesting, since the time between sampling and taxonomic identification is reduced. Due to the presence of non-target material like plants and sediments in crude samples, DNA extraction protocols become important for maximising DNA recovery and sample replicability. We sampled freshwater invertebrates from six river and lake sites and extracted DNA from homogenised bulk samples in quadruplicate subsamples, using a published method and two commercially available kits: HotSHOT approach, Qiagen DNeasy Blood & Tissue Kit and Qiagen DNeasy PowerPlant Pro Kit. The performance of the selected extraction methods was evaluated by measuring DNA yield and applying DNA metabarcoding to see if the choice of DNA extraction method affects DNA yield and metazoan diversity results. The PowerPlant Kit extractions resulted in the highest DNA yield and a strong significant correlation between sample weight and DNA yield, while the DNA yields of the Blood & Tissue Kit and HotSHOT method did not correlate with the sample weights. Metazoan diversity measures were more repeatable in samples extracted with the PowerPlant Kit compared to those extracted with the HotSHOT method or the Blood & Tissue Kit. Subsampling using Blood & Tissue Kit and HotSHOT extraction failed to describe the same community in the lake samples. Our study exemplifies that the choice of DNA extraction protocol influences the DNA yield as well as the subsequent community analysis. Based on our results, low specimen abundance samples will likely provide more stable results if specimens are sorted prior to DNA extraction and DNA metabarcoding, but the repeatability of the DNA extraction and DNA metabarcoding results was close to ideal in high specimen abundance samples.

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Simple, rapid and cost-effective DNA extraction method for high quality DNA suitable for PCR based downstream application in mungbean [Vigna radiata (L.) Wilczek]

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/346/1/012037 ◽

2019 ◽

Vol 346 ◽

pp. 012037

Author(s):

P Pookhamsak ◽

P Pornbungkerd ◽

P A Tantasawat

Keyword(s):

Dna Extraction ◽

Vigna Radiata ◽

Extraction Method ◽

Cost Effective ◽

High Quality ◽

Dna Extraction Method ◽

Downstream Application

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Sensitivity detection of Abacavir in human through SNP detection of HLA-B*5701 allele

International Journal of Bioassays ◽

10.21746/ijbio.2016.08.006 ◽

2016 ◽

Vol 5 (08) ◽

pp. 4754

Author(s):

Tanushree Mitra* ◽

Shivshankar Kumdale ◽

Sameer Chowdhary ◽

Amol D. Raut

Keyword(s):

Blood Sample ◽

Dna Extraction ◽

Genomic Dna ◽

Extraction Method ◽

Snp Detection ◽

Dna Extraction Method ◽

Sensitivity Detection

The main objective of this study was to make sure whether randomly taken 12 samples were sensitive to abacavir. The genomic DNA from 12 blood sample were extracted by phenol chloroform DNA extraction method, extracted genomic DNA were amplified and sequenced, thereafter SNPs were detected. Every sample had shown the presence of normal base at SNP position. This study indicated, those randomly taken 12 patients were sensitive to abacavir, so they can consume abacavir if they get infected with HIV.

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