ISOLATE
RNA Kits
7
R
N
A
M
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i K
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6. equipment anD reagents to Be supplieD By the user
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Suitable container to hold sample
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Mortar and pestle and liquid N
2
or rotor stator homogenizer
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Microcentrifuge with rotor for 1.5ml and 2.0ml tubes
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Shaking platform
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70% and 96-100% ethanol
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TE Buffer
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ddH
2
O
7. protocols
7.1 total rna isolation from animal tissue
Before you start:
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Before using for the first time, add 96-100% ethanol to the Wash Buffers AR
and BR as indicated on the bottles and mix.
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If any of the buffers form precipitates upon storage, re-dissolve by gently
warming. Cool to room temperature before use.
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If using frozen tissue, do not allow tissue to thaw during weighing or before the
addition of Lysis Buffer R. Once homogenized in the Lysis Buffer, the sample
can be stored at -20ºC for several months.
•
For information on how to work with RNA, read Hints and Tips on page 19.
1. Homogenize and lyse up to 20mg of tissue sample using liquid nitrogen or
a rotor-stator homogenizer.
Using liquid nitrogen
1.1. Grind the sample to a fine powder using a mortar and pestle in the
presence of liquid nitrogen. Take care that the sample does not thaw during or
after grinding.
1.2. Transfer the sample to a 1.5ml microcentrifuge tube (not supplied).
1.3. Immediately add 450µl Lysis Buffer R and homogenize the sample.
Proceed to next step. The sample can also be stored at this step at -20ºC.
Using a rotor-stator homogenizer
1.1 Transfer the sample to a suitable container.
1.2 Add 450µl Lysis Buffer R and homogenize the sample.
1.3 Transfer the sample to a 1.5ml microcentrifuge tube (not supplied).
Proceed to next step. The sample can also be stored at this step at -20ºC.
