peqGOLD HP Total RNA Kit - PEQLAB Biotechnologie GmbH

TROUBLESHOOTING TIPS
Problem Problem
Likely Likely cause cause
Suggestion
Suggestion
Repeat elution.
Pre-heat RNase-free Water to 70 o Little or no RNA
eluted
RNA remains on the column
C prior to
elution.
Incubate column for 10 min with RNase-free
Water prior to centrifugation.
Column is overloaded Reduce quantity of starting material.
Clogged column Incomplete homogenization
Degraded RNA
Problem in
downstream
applications
DNA
contamination
Low Absorption
ratios
Source
RNase contamination
Salt carry-over during
elution
Completely homogenize sample.
Increase centrifugation time.
Reduce amount of starting material
Freeze starting material quickly in liquid
nitrogen.
Do not store tissue culture cells prior to
extraction unless they are lysed first.
Follow protocol closely, and work quickly.
Ensure not to introduce RNase during the
procedure.
Check buffers for RNase contamination.
Ensure RNA Wash Buffer II Concentrate has
been diluted with 4 volumes of 100 % ethanol
as indicated on the bottle.
RNA Wash Buffer II must be stored and used
at room temperature.
Repeat wash with RNAWash Buffer II.
Co-purification of DNA Digest with RNase-free DNase and incubate
at 37 o C for 5 min.
RNA diluted in acidic buffer
or water
RNase-free Water is acidic and can
dramatically lower Abs 260 values. Use TE
buffer to dilute RNA prior to
spectrophotometric analysis.
peqGOLD HP Total RNA Kit Ord.-No.: 12-6935/12-6936
V.0111 PEQLAB Biotechnologie GmbH 18