Follow
International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39 For more details click here
National Academy of Agricultural Sciences (NAAS) : NAAS Score: *5.38 (2020) [Effective from January 1, 2020]For more details click here

Login as a Reviewer

Indexed in



National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2018: 95.39
For more details click here

See Guidelines to Authors
Current Issues

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 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2020.9(4): 1753-1756
DOI: https://doi.org/10.20546/ijcmas.2020.904.205


An Easy Method to Isolate the High Quality DNA from Plant Pathogen Alternaria spp
Sushma Nema, Keerti Tantwai* and Lalit Prasad Singh Rajput
Biotechnology Centre, Jawaharlal Nehru Krishi Vishwa Vidyalaya, Jabalpur – 482004, MP, India
*Corresponding author
Abstract:

A simple and easy protocol for isolation of DNA from major plant pathogen Alternaria spp has been developed in which DNA extracted using CTAB extraction method from fresh mycelium of fungi crushed with sterile sand particles and liquid nitrogen. The DNA was precipitated first with equal volume of phenol-chloroform-isoamylalcohol (25:24:1) and further with equal volume of chloroform-isoamylalcohol (24:1). Repeated centrifugation was done at 10000 g for 10 minutes at room temperature. DNA was pooled by incubating the supernatant with pre-chilled isopropanol at -20°C for 1 hour. After that the samples were centrifuged for 15 minutes at 13000 g to pellet the DNA. The DNA pellet was washed with 200 µL of 70% ethanol, air dried and dissolved in 200 µL of TE buffer, thereafter 5 µL of RNase was added to each DNA Samples, mixed and incubated at 37°C for 1 hour. After RNase treatment, DNA samples were precipitated with chloroform-isoamylalcohol (24:1) again incubated with pre-chilled isopropanol and then DNA pellet which was finally dissolved in 20 µL of TE buffer and stored at -20°C till further use. Inter-simple sequence repeats (ISSRs) amplification reaction was carried out using 20 µL of reaction volume containing 2 µL of DNA.


Keywords: Alternaria, ISSR, DNA isolation, molecular marker
Download this article as Download

How to cite this article:

Sushma Nema, Keerti Tantwai and Lalit Prasad Singh Rajput. 2020. An Easy Method to Isolate the High Quality DNA from Plant Pathogen Alternaria Spp.Int.J.Curr.Microbiol.App.Sci. 9(4): 1753-1756. doi: https://doi.org/10.20546/ijcmas.2020.904.205