Bio-Rad PureZOL™ RNA Isolation Reagent User Manual
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Section 6
Troubleshooting
Problem
Possible Cause
Recommended Solution
Incomplete
Lysate was not mixed
Once chloroform is added,
separation of
properly after adding
mix tubes vigorously by
phases after
chloroform (See step 3
shaking for 15 seconds. Do
centrifugation
in protocol)
not vortex. Let the lysate
incubate for 5 minutes at
room temperature, then mix
again before centrifuging
Lysate was not
Make sure that the
centrifuged at the right
centrifugation is performed
temperature
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
Low yield
Incomplete disruption
Increase the duration or
and homogenization
intensity of sample
of sample
disrup tion.Scale up the
amount of PureZOL used
based on sample size
Over dried RNA pellet
Do not use a Speed Vac to
dry the pellet. Dry pellet
briefly at room temperature
for 5 minutes. Add DEPC-
treated water to the tube
and pipet up and down
several times to resuspend
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