Bio-Rad Aurum™ Total RNA 96 Kit User Manual

Page 20

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