METHOD FOR ISOLATING DNA FROM PLANT PATHOGENS

Abstract

ABSTRACT A method (100) for isolating DNA from plant pathogens, the method (100) comprising the steps of collecting plant tissue samples from field-grown plants infected by pathogens, crushing the plant tissue samples into a fine powder and combining them with a cetyltrimethylammonium bromide (CTAB) extraction buffer to obtain a mixture, incubating the mixture at a temperature of 65°C for a specific time duration, adding a chloroform-isoamyl alcohol solution to the mixture to separate the DNA-containing aqueous phase, precipitating DNA from the DNA-containing aqueous phase using cold isopropanol, followed by centrifugation to obtain a DNA pellet and washing and drying the DNA pellet and suspending the DNA pellet in a suitable buffer to obtain a purified DNA.

Specification

Description:FIELD OF THE DISCLOSURE
[0001] This invention generally relates to a field of DNA isolation and in particular relates to a method for isolating DNA from plant pathogens.
BACKGROUND

[0002] The subject matter discussed in the background section should not be assumed to be prior art merely as a result of its mention in the background section. Similarly, a problem mentioned in the background section or associated with the subject matter of the background section should not be assumed to have been previously recognized in the prior art. The subject matter in the background section merely represents different approaches, which in and of themselves may also correspond to implementations of the claimed technology.
[0003] The presence of plant pathogens, including viruses, bacteria, fungi, and nematodes, poses a significant threat to crop yield and agricultural sustainability worldwide. Early detection and identification of these pathogens are critical for effective disease management and prevention of widespread , Claims:1. A method (100) for isolating DNA from plant pathogens, the method (100) comprising the steps of:
collecting plant tissue samples from field-grown plants infected by pathogens;
crushing the plant tissue samples into a fine powder and combining them with a cetyltrimethylammonium bromide (CTAB) extraction buffer to obtain a mixture;
incubating the mixture at a temperature of 65°C for a specific time duration;
adding a chloroform-isoamyl alcohol solution to the mixture to separate the DNA-containing aqueous phase;
precipitating DNA from the DNA-containing aqueous phase using cold isopropanol, followed by centrifugation to obtain a DNA pellet; and
washing and drying the DNA pellet and suspending the DNA pellet in a suitable buffer to obtain a purified DNA.

2. The method (100) as claimed in claim 1, wherein the CTAB extraction buffer is prepared with a pH of 8.0 and includes Tris-HCl, Na2EDTA, and NaCl to enhance DNA purity and yield.

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