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P R O T O C O L S The
comet assay and microgel electrophoresis (MGE) were first introduced by Ostling
and Johanson in 1984. This was a neutral assay in which the lysis and
electrophoresis were done under neutral conditions. Staining
was done with acridine orange. The image obtained looked like a “Comet?with
a distinct head, comprising of intact DNA and a tail, consisting of damaged or
broken pieces of DNA hence the name “Comet?Assay was given. The extent of
DNA liberated from the head of the comet was the function of the dose of
irradiation. However, in this procedure, only double strand breaks could be
analyzed. The assay
can be performed on a variety of samples which can be obtained as a single
cell population e.g. peripheral blood lymphocytes, nasal and buccal
epithelium from clinically or occupationally exposed human population and
for in vitro studies on cell
lines e.g. CHO, V79, mouse lymphoma or cultured human lymphocytes and bone
marrow cells. Both DNA damage and repair studies can be conducted. Also a
variety of information related to genetic toxicology, human epidemiology,
patients undergoing radio/chemo- therapy, ageing and nutrition can be obtained. The
assay has been used for environmental biomonitoring and has utilized
earthworms, fishes, mollusks exposed to polluted environments.
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DETAILED PROTOCOLS FOR COMET ASSAY AND FOR APOPTOSIS
COMET
ASSAY PROTOCOL PROTOCOL FOR DNA STRAND
BREAKS PROTOCOL
FOR COMET ASSAY IN SPERM *THESE PDF FILES ARE ONLY FOR PERSONAL USE AND NOT FOR DISTRIBUTION OR COMMERCIAL PURPOSES
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