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A method of direct PCR without DNA extraction for rapid detection of begomoviruses infecting jute and mesta

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Title A method of direct PCR without DNA extraction for rapid detection of begomoviruses infecting jute and mesta
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Creator C. Biswas, P. Dey and S. Satpathy
 
Subject direct PCR, jute, lysis buffer, mesta.
 
Description Not Available
We have developed a simple method of direct PCR (dPCR) without timeconsuming procedures of DNA extraction by directly using the leaf bits for
rapid detection of begomoviruses in jute and mesta. The leaf bits were treated
with a lysis buffer for 35 min, and the lysate was used as PCR template.
Different components and their concentration in lysis buffer systems were
optimized and the optimal buffer system composed of 20 mmol l 1 tris
(hydroxymethyl aminomethane (Tris)-Cl (pH 80)), 15 mmol l 1
ethylenediaminetetraacetic acid (EDTA) (pH 80), 14 mol l 1 NaCl and
200 lg/mL Proteinase K. Further, 3% PVP (w/v) and b-marcaptoethanol (1%
v/v) were additionally added into the buffer in case of jute. Under optimized
PCR conditions, both viral DNA as well as plant (jute and mesta) genomic
DNA were amplified from the lysate. dPCR required fewer reagents and less
incubation time reducing both time and cost of detection.
Not Available
 
Date 2020-08-06T08:38:12Z
2020-08-06T08:38:12Z
2013-11-20
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/38999
 
Language English
 
Relation Not Available;
 
Publisher Wiley-Blackwell Publishing Ltd