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Evaluation of different methods of DNA extraction from semen of buffalo (Bubalus bubalis) bulls

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Title Evaluation of different methods of DNA extraction from semen of buffalo (Bubalus bubalis) bulls
Not Available
 
Creator Anju Manuja
Sonia Manchanda
Balvinder Kumar
S Khanna
RK Sethi
 
Subject DNA extraction, PCR inhibitors, semen, Bubalus bubalis, buffalo bull
 
Description Not Available
Microbes excreted in the semen of infected
or carrier bulls can be disseminated to susceptible
animals through artificial insemination. The
polymerase chain reaction (PCR) has been
employed successfully to detect infectious agents
in tissues and body fluids. PCR inhibitors present in
the semen pose serious problems in detection of
microorganisms by inhibiting the amplification of the
target DNA template. These inhibitors need to be
removed completely during DNA extraction to
amplify the target sequences in semen by PCR.
DNA was extracted using seven different protocols
from semen of buffalo bulls, and the quantity and
quality was evaluated spectrophotometrically.
Chelex-100 and Qiagen modified methods for
extraction of DNA from semen were found to be
superior qualitatively as compared to the other
methods. In the Qiagen modified protocol, the semen
was treated with two extra buffers containing EDTA
to chelate the metals. Additional treatment of semen
with proteinase K was included to completely
degrade cellular proteins. DNA extracted by the
phenol-chloroform and CTAB methods yielded high
value of residual RNA and other contaminants. The
chelex-100 method has the potential advantage of
requiring a smaller volume of semen to extract good
quality of DNA.
Not Available
 
Date 2022-09-29T09:46:07Z
2022-09-29T09:46:07Z
2010-06-01
 
Type Article
 
Identifier Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/74609
 
Language English
 
Relation Not Available;
 
Publisher Not Available