Total RNA, DNA, and
protein isolation from brain tissue were performed using the NucleoSpinTriPrep
(Macherey-Nagel) mini kit for RNA, DNA, and protein purification.
Thus, it ensured the parallel isolation of RNA, DNA, and protein from
an undivided sample at a higher yield and purity.46 (link) For this purpose, the left hemispheres of mice brains were
ground to fine powders in the presence of liquid nitrogen using a
DEPC-treated mortar and pestle. Then cells are lysed by incubation
in a solution containing large amounts of chaotropic ions and applied
to the Triprep column. DNA, RNA, and protein were isolated separately
and sequentially according to the manufacturer’s instructions.
The obtained RNA and DNA quality were visualized in agarose gel electrophoresis,
and their concentrations were measured in the “NanoDrop”
device. After RNA isolation, cDNA synthesis was performed immediately
from the RNAs whose concentrations were calculated.
protein isolation from brain tissue were performed using the NucleoSpinTriPrep
(Macherey-Nagel) mini kit for RNA, DNA, and protein purification.
Thus, it ensured the parallel isolation of RNA, DNA, and protein from
an undivided sample at a higher yield and purity.46 (link) For this purpose, the left hemispheres of mice brains were
ground to fine powders in the presence of liquid nitrogen using a
DEPC-treated mortar and pestle. Then cells are lysed by incubation
in a solution containing large amounts of chaotropic ions and applied
to the Triprep column. DNA, RNA, and protein were isolated separately
and sequentially according to the manufacturer’s instructions.
The obtained RNA and DNA quality were visualized in agarose gel electrophoresis,
and their concentrations were measured in the “NanoDrop”
device. After RNA isolation, cDNA synthesis was performed immediately
from the RNAs whose concentrations were calculated.
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