Comparative analysis of Genomic DNA extraction protocols: Maxi-Preparation of Quality DNA for  Genetic Evaluation

Naeem, Zara; Masud, Samra; Hassan, Sk. Sarif; Naeem, M.

doi:10.26692/surj/2020.03.02

Cited by 3 publications

(2 citation statements)

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“…Then DNA pellets were washed with 70% alcohol (chilled). 60 µl of nuclease free water was added to re-suspend the DNA [11].…”

Section: Pipetted Then Lysate Was Transferred Into Genejetmentioning

confidence: 99%

“…DNA isolation. Six traditional DNA isolation techniques whose methodologies had been described previously for fishes were selected from the literature and conditions of DNA isolation were intended to remain same for TNES [9], Phenol chloroform [10], Urea SDS [11], SNET [12], Rapid MT [13] and Salt out methods [12]. Seventh proposed, evaluated and compared method was a commercially available GeneJET Genomic DNA Purification Kit.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of seven genomic DNA isolation techniques from internal organs, gills and muscle tissues of Notopterus notopterus (Notopteridae) using PCR amplification and Nanodrop

Ahmad

2023

IJBCh

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The techniques and the principles used in DNA isolation play a vital role in the obtaining of a purified genetic material. In present study, we investigated the efficiency of seven genomic DNA isolation techniques in terms of isolated DNA concentration, yield and purity from internal organs, gills and muscle tissues of Notopterus notopterus. Isolated DNA quality was analysed through Nanodrod and PCR using mitochondrial COI genetic markers. Results showed that GeneJET Genomic DNA Purification Kit was found significantly higher in terms of isolated DNA concentration (1200-1288 ng.ul -1 ), yield (257.6 ng.ul -1 ) and purity (1.91-2.00), and also successful in PCR amplification as compared to other evaluated six traditional DNA isolation methods. The nucleotide minimum bands range was observed 200 base pairs in heart sample and maximum 600 base pair was observed in intestine. There is no data on description of parameters analysed in this work to date, neither the evaluation of isolated DNA using PCR amplification of mitochondrial COI gene for the species N. notopterus. Present study also revealed that the traditional DNA isolation methods are the secondary choice for isolation of DNA. The data of present study also indicated that the GeneJET Genomic DNA Purification Kit is useful for DNA isolation and can be used best in genetic applications for fishes.

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“…Then DNA pellets were washed with 70% alcohol (chilled). 60 µl of nuclease free water was added to re-suspend the DNA [11].…”

Section: Pipetted Then Lysate Was Transferred Into Genejetmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of seven genomic DNA isolation techniques from internal organs, gills and muscle tissues of Notopterus notopterus (Notopteridae) using PCR amplification and Nanodrop

Ahmad

2023

IJBCh

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An efficient, fast and inexpensive method for genomic DNA extraction of fish tissue

2023

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Comparative Analysis of Dna Isolated From Fish Samples by Three Different Extraction Protocols

Ot¹,

Biotechnology²,

Da³

et al. 2020

JEBAS

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The studies on fish genetic diversity and its conservation in Nigeria are still at its preliminary stages. The country needs to document the diversities of all the water bodies and also embark on a DNA barcoding project for rapid identification of the enormous populations and consequent deposition in the global genebank for references. All molecular studies usually start with the isolation, purification, and recovery of DNA and this usually depends on the types of tissue, mode of sample collection, the medium of storage, duration of storage, and used extraction protocols. The current study embarked on fish collection in four major freshwater habitats as a preliminary study to a proposed fish barcoding project and to comparatively determine the extraction protocol that will be cost-effective, fast, safe, and yield adequate molecular materials for downstream amplification, cloning, and sequencing reactions. In the current study, three DNA extraction protocols, Zymo Research Kit (ZR), modified conventional SNET method, and modified Urea-SDS Method were compared to establish the best DNA extraction method from freshwater fishes. Sixty-two (62) fish samples were collected belonging to 16 different families, 23 Genera, and 32 Species. The average yield of the three protocols in terms of concentration (ng/μL: Purity) are: ZR (30.59: 1.58); UREA, (705.49: 1.75) and SNET (562.22: 1.73). Hence, in terms of DNA concentration recovery, the sequence of the best method is UREA > SNET > ZR, and the same trend followed in the case of Purity. Statistical tests did not show any significant difference when the extraction protocols were compared among fish families. Cytochrome B gene was successfully amplified on the DNA template to confirm their suitability for further studies. The result of the study can be concluded that among the best DNA extraction methods, UREA protocol can be recommended for fish DNA extraction, this is not only cost-effective, but also gave quality yield and adequate for downstream analysis.

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Comparative analysis of Genomic DNA extraction protocols: Maxi-Preparation of Quality DNA for Genetic Evaluation

Cited by 3 publications

References 0 publications

Comparison of seven genomic DNA isolation techniques from internal organs, gills and muscle tissues of Notopterus notopterus (Notopteridae) using PCR amplification and Nanodrop

Comparison of seven genomic DNA isolation techniques from internal organs, gills and muscle tissues of Notopterus notopterus (Notopteridae) using PCR amplification and Nanodrop

An efficient, fast and inexpensive method for genomic DNA extraction of fish tissue

Comparative Analysis of Dna Isolated From Fish Samples by Three Different Extraction Protocols

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