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192798

Genomic DNA extraction from minimal amount of dried mushroom samples

Article

Last updated: 23 Dec 2024

Subjects

-

Tags

Agricultural Microbiology
Biodiversity and Conservation
Ecosystem ecology
Fungi
Microbial-plant interactions
Plant Pathology
Symbiosis

Abstract

Aims: To develop a simple and rapid genomic DNA extraction technique for dried (≈ 1 year old) mushroom fruiting bodies that yields high-quality DNA, suitable for use by post-graduate institutions. Method: Small amounts (0.04 g) of pulverized dried mushroom sample were incubated in a Tris /EDTA/SDS lysis buffer (100mM:10mM: 2%) at 65°C to lyse the chitinous fungal cell walls. Genomic DNA purification was performed using chloroform isoamyl alcohol (24:1), and DNA was precipitated using 100% ethanol. Results: Genomic DNA was successfully extracted under 70 minutes from 16 samples morphologically identified as Panaeolus, Copelandia, Gymnopilus, Pluteus and Favolus species. DNA concentrations were on average of 696.9ng/µL. PCR successfully amplified the ITS-5.8S region.  The protocol has been successfully used by numerous post-graduate students in our research programme. Conclusion: The rapid and easy protocol produced high-quality genomic DNA void of any inhibitors that is suitable for downstream molecular implications across multiple mushroom genera.  Noticeably, this method requires only minute quantities (0.04g) of starting material and is ideal for student training in higher academic institutions.

DOI

10.21608/mb.2021.91637.1036

Keywords

Affordable, DNA extraction, dried mushroom specimens, rapid, student friendly

Authors

First Name

Dominique

Last Name

Strauss

MiddleName

-

Affiliation

Department of Genetics, Natural and Agricultural Sciences, University of Free State, 205 Nelson Mandela Dr, Park West, Bloemfontein, 9301, South Africa

Email

research.strauss@gmail.com

City

Bloemfontein

Orcid

-

First Name

Soumya

Last Name

Ghosh

MiddleName

-

Affiliation

Department of Genetics, Natural and Agricultural Sciences, University of Free State, 205 Nelson Mandela Dr, Park West, Bloemfontein, 9301, South Africa

Email

soumyaghosh@yahoo.com

City

Bloemfontein

Orcid

0000-0002-4945-3516

First Name

Zurika

Last Name

Murray

MiddleName

-

Affiliation

Department of Genetics, Natural and Agricultural Sciences, University of Free State, 205 Nelson Mandela Dr, Park West, Bloemfontein, 9301, South Africa

Email

murrayz@ufs.ac.za

City

-

Orcid

-

First Name

Marieka

Last Name

Gryzenhout

MiddleName

-

Affiliation

Department of Genetics, Natural and Agricultural Sciences, University of Free State, 205 Nelson Mandela Dr, Park West, Bloemfontein, 9301, South Africa

Email

gryzenhoutm@ufs.ac.za

City

-

Orcid

-

Volume

6

Article Issue

1

Related Issue

24927

Issue Date

2021-06-01

Receive Date

2021-08-19

Publish Date

2021-09-04

Page Start

49

Page End

54

Print ISSN

2357-0326

Online ISSN

2357-0334

Link

https://mb.journals.ekb.eg/article_192798.html

Detail API

https://mb.journals.ekb.eg/service?article_code=192798

Order

6

Type

Original Article

Type Code

502

Publication Type

Journal

Publication Title

Microbial Biosystems

Publication Link

https://mb.journals.ekb.eg/

MainTitle

Genomic DNA extraction from minimal amount of dried mushroom samples

Details

Type

Article

Created At

22 Jan 2023