Bio-Rad PureZOL™ RNA Isolation Reagent - Manual (Page 14)

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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