Bio-Rad Aurum™ Total RNA 96 Kit - Manual (Page 21)

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Problem Possible Cause Recommended Solution
Clogging of RNA Excessive amount of Reduce volume of
binding plate starting material per well culture used
Incomplete mixing of lysis Mix RNA lysis solution
and alcohol solutions and alcohol thoroughly
by pipetting up and
down
Incomplete Pipette lysate up and
homogenization of cell down further to reduce
lysate lysate viscosity
Incomplete digestion with Increase duration of
lysozyme or lyticase lysozyme or lyticase
digestion; use fresh
enzyme
Low RNA yield Low amount of starting Increase starting material
material amount up to the
maximum indicated for
the specific starting
material type
Incorrect use of wash Add the appropriate
solutions volume of 95–100%
ethanol to the low
stringency wash solution
before initial use
Incorrect preparation Use only the DNase
of DNase dilution dilution solution provided
in the kit to dilute the
DNase I
Low sample eluate See problem “Low or
volume highly variable eluate
volumes among wells”
Inefficient elution Preheat the elution
solution to 70°C in water
bath prior to the elution
step
Incomplete lysis Pipet lysate up and down
further to facilitate lysis