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An efficient method of mitochondrial DNA isolation from Vigna radiata for genomic studies

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Title An efficient method of mitochondrial DNA isolation from Vigna radiata for genomic studies
 
Creator Singh, Pooja
Sahoo, Ranjan Kumar
Bulle, Mallesham
Gupta, Kapuganti Jagadis
 
Subject β-Actin
DNA
Mitochondria
Mung bean
Percoll gradient
 
Description Accepted date: 1 Jan 2020
Isolation of mitochondrial DNA from root tissues of mung bean (Vigna radiata) is quite tedious, complex, and often results in low yield. Hence here we show a simple, rapid, and improved protocol for isolation of mitochondrial DNA from root tissues of hydroponically grown mung bean plants. This method involves purification of mitochondria and subsequent isolation of DNA from obtained purified mitochondria. For this purpose, mitochondria were isolated using a discontinuous Percoll gradient centrifugation followed by RNase I treatment to the isolated DNA to remove any traces of RNA contamination. The mitochondrial DNA was isolated from mitochondrial samples by commonly used CTAB method. The specificity of isolated mitochondrial DNA was confirmed using mtDNA-specific genes (NAD1 and COX3). β-Actin primer was used to check the nuclear DNA contamination. PCR amplification was observed in mtDNA specific genes NAD1 and COX3 except nuclear encoded β-actin gene suggesting that mitochondrial DNA was not contaminated by nuclear DNA.
PS is supported by a senior research fellowship from UGC, India.
 
Date 2020-01-07T06:39:23Z
2020-01-07T06:39:23Z
2020
 
Type Article
 
Identifier Methods in Molecular Biology, 2107: 305-315
978-1-0716-0235-5
https://doi.org/10.1007/978-1-0716-0235-5_16
https://link.springer.com/protocol/10.1007%2F978-1-0716-0235-5_16
http://223.31.159.10:8080/jspui/handle/123456789/1030
 
Language en_US
 
Format application/pdf
 
Publisher Springer Nature Publishing AG