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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(3): 1619-1624
DOI: https://doi.org/10.20546/ijcmas.2019.803.188


A Simple and Efficient Genomic DNA Extraction Protocol for Dried Leaf of Threatened Species Commiphora wightii (Arnott) Bhandari for Genetic Analysis of Plant Biological System
Sajjan Kumar Pooniya1*, Keerti Tantwai1, Niraj Triphati1 and Sharad Tiwari1,2
1Biotechnology Centre
2Department of Plant Breeding and Genetics, Jawaharlal Nehru Krishi Vishwa Vidyalaya, Jabalpur, Madhya Pradesh, 482004, India
*Corresponding author
Abstract:

Commiphora wightii (Arnott) Bhandari contains secondary metabolites, polysaccharides and phenolic compounds. The presence of secondary metabolites, reduce the yield and quality of the DNA. In the present study an alternative protocol for genomic DNA extraction from dry plant leaves was developed that is acquiescent to PCR-based genomic studies. Existing protocols were lengthy, costly or not appropriate for genomic DNA extraction from dry leaves. This modified CTAB (3%) and PVP (Polyvinylpyrrolidone) 1.5% protocol include the use of 0.5M NaCl, 0.3% ß-mercaptoethanol in the extraction as well as application of autoclaved sand for proper grinding of dried leaves and inclusion of RNase A treatment in the protocol to fasten the process. The extracted DNA using present optimized protocol was super in quality and quantity. It was also suitable for polymerase chain reaction with random decamer, inter simple sequence repeat and barcode primers. The developed protocol is rapid and cost efficient with high quality and sufficient quantity of DNA for downstream PCR-based genetic analysis.


Keywords: Commiphora wightii, Molecular markers, PCR amplification, polyvinyl pyrrolidone, ß-mercaptoethanol, DNA extraction
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How to cite this article:

Sajjan Kumar Pooniya, Keerti Tantwai, Niraj Triphati and Sharad Tiwari. 2019. A Simple and Efficient Genomic DNA Extraction Protocol for Dried Leaf of Threatened Species Commiphora wightii (Arnott) Bhandari for Genetic Analysis of Plant Biological System.Int.J.Curr.Microbiol.App.Sci. 8(3): 1619-1624. doi: https://doi.org/10.20546/ijcmas.2019.803.188