Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit - Manual (Page 32)

Problem Possible Cause Recommended Solution
Genomic DNA On-column DNase I Perform the on-column
contamination digest was not DNase I digest (see step
performed 14 in protocol)
Incomplete DNase I Increase the digest time
digest for starting materials that
are known to contain a
high level of genomic
DNA
DNase I is inactive Reconstitute the
lyophilized DNase I with
10 mM Tris, pH 7.5.
Aliquot and store the
reconstituted DNase I
enzyme in a nonfrost-free
freezer. Avoid freeze-
thaw cycles by aliquoting
the enzyme for single use
only
Some of the white Leave some of the
interphase (after aqueous phase solution
phase separation) behind to avoid
was transferred with transferring the white
the aqueous phase interphase with the
aqueous phase (see
step 7 in the protocol)
RNA is degraded RNase contamination Make sure to use
of solutions provided RNase-free plasticware.
in the kit See Section 5,
Maintaining an
RNase-free Environment,
for detailed instructions
RNase contamination Treat all user-made
of solutions supplied solutions with DEPC
by the user before use (see Section 6
for instructions)
RNase contamination See Section 6,
of plasticware and Maintaining an RNase-free
work station Environment, for detailed
instructions
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