Adults of each insect species were individually
ground in 400 μl of cold extraction buffer (10 mM Tris, pH 8.0, 0.1 M
EDTA, 20 μg/ml RNase and 0.5% SDS)
The ground sample was
incubated at 50 °C for 10 min with 5 μl of proteinase K (20 mg/ml).
To remove macromolecules other than nucleic acids, a mixture of
phenol (pH 8.0) and chloroform (1:1, v/v) was added in a volume
equal to the bground?N sample.The mixture was then centrifuged at
14,000 rpm for 2 min. The upper layer was removed and mixed with
an equal volume of a mixture of chloroform and isopropyl alcohol
(24:1, v/v). Genomic DNA was then isolated by ethanol precipitation
and resuspended in sterilized deionized water
Adults of each insect species were individually
ground in 400 μl of cold extraction buffer (10 mM Tris, pH 8.0, 0.1 M
EDTA, 20 μg/ml RNase and 0.5% SDS)
The ground sample was
incubated at 50 °C for 10 min with 5 μl of proteinase K (20 mg/ml).
To remove macromolecules other than nucleic acids, a mixture of
phenol (pH 8.0) and chloroform (1:1, v/v) was added in a volume
equal to the bground?N sample.The mixture was then centrifuged at
14,000 rpm for 2 min. The upper layer was removed and mixed with
an equal volume of a mixture of chloroform and isopropyl alcohol
(24:1, v/v). Genomic DNA was then isolated by ethanol precipitation
and resuspended in sterilized deionized water
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