Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit - Manual (Page 29)

Section 9
Troubleshooting Guide
Problems that may be encountered during RNA purification:
Problem Possible Cause Recommended Solution
Incomplete Lysate was not mixed Once chloroform is
separation of properly after adding added, mix tubes
phases after chloroform (see step 4 vigorously by shaking
centrifugation in protocol) for 15 sec. Do not vortex!
Let the lysate incubate for
5 min at room
temperature, mix again
before centrifuging
Lysate was not Make sure that
centrifuged at the right centrifugation step is
temperature performed at 4°C
following the addition of
chloroform in order to
achieve complete
separation of the phases
Incorrect amount of For every 1 ml of PureZOL™
chloroform was added used, add 0.2 ml of
chloroform
RNA binding mini Incomplete disruption of Increase the duration or
column is clogging starting material intensity of sample
disruption. Make sure to
use the 1 ml of PureZOL
for each prep
Excessive amount of Do not exceed the
starting material maximum starting amount
limit for the kit (see
Section 5). If clogging
persists when using the
maximum starting
amount, reduce the
amount of material used
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