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Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit User Manual

Page 29

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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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