Bio-Rad Aurum™ Total RNA 96 Kit - Manual (Page 20)

16
Problem Possible Cause Recommended Solution
Low or highly variable Elution solution applied Apply elution solution
eluate volumes among to RNA binding plate well directly to membranes at
wells
walls base of each well
RNA binding plate not Set plate properly and
seated properly press down gently to
seat
Abrupt application of Increase negative
negative pressure during pressure gradually
elution over 5–10 sec using
the vacuum regulator
Residual wash buffer on Blot RNA binding plate
drip directors drip directors with paper
towels; ensure that the
plate underside is dry
Genomic DNA Incomplete DNase I Increase DNase I digestion
contamination digestion time
Inactive DNase I Store reconstituted
DNase I in a nonfrost-
free freezer; avoid
freeze-thaw cycles;
aliquot reconstituted
DNase I for single use
only
Excessive amount of Reduce volume of
starting material per well culture used
Incorrect preparation Use only the DNase
of DNase dilution dilution solution provided
in the kit to dilute the
DNase
RNA degradation RNase contamination of DEPC-treat all handmade
user-made solutions solutions; decontaminate
and/or plasticware all work surfaces; see
Section 5 for more
details
Endogenous RNases Work quickly through
the steps prior to the
addition of lysis solution