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12
RNeasy Mini Handbook 10/2019
Description of protocols
Purification of Total RNA from Animal Cells Using Spin Technology
Up to 1 x 10
7
cells, depending on the cell line, are disrupted in Buffer RLT and homogenized.
An overview of disruption and homogenization methods is given on page 22. Ethanol is then
added to the lysate, creating conditions that promote selective binding of RNA to the RNeasy
membrane. The sample is then applied to the RNeasy Mini spin column. Total RNA binds to
the membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in
RNAse-free water. All bind, wash, and elution steps are performed by centrifugation in a
microcentrifuge.
Purification of Total RNA from Animal Cells Using Vacuum/Spin Technology
Up to 1 x 10
6
cells, depending on the cell line, are disrupted in Buffer RLT and homogenized.
An overview of disruption and homogenization methods is given on page 22. Ethanol is then
added to the lysate, creating conditions that promote selective binding of RNA to the RNeasy
membrane. The sample is then applied to the RNeasy Mini spin column. Total RNA binds to
the membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in
RNAse-free water. The bind and wash steps are performed on a QIAvac 24, QIAvac 24 Plus,
or QIAvac 6S vacuum manifold, and the final elution step is performed by centrifugation in a
microcentrifuge.
Stabilization of RNA in Harvested Animal Tissues
This protocol describes how to stabilize RNA in harvested animal tissues using RNAprotect
Tissue Reagent. Purification of total RNA from the stabilized tissues can be subsequently carried
out according to “Protocol: Purification of Total RNA from Animal Tissues” (page 45).
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