The following tables contain the compounds tested and indicate the compatibility of the compound tested with RNase A activity.

Experimental procedure: 45 µl of each listed reagent was added to an RNaseAlert® tube containing lyophilized substrate plus 5 µl of 10X RNaseAlert® Reaction Buffer. These mixtures were incubated for 1 hr at 37°C and then inspected for fluorescence on a UV transilluminator. Sample fluorescence was marked "-" in the "Before Addition of RNase A" if there was no visible glow and "+" if there was visible green glow.

After UV inspection and scoring, the "-" samples were further tested by adding 100 pg of RNase A and incubating an additional hour at 37°C. These tubes were then scored as above and results listed in the "After Addition of RNase A" column. RNase A was not added to enzyme solutions possessing RNase activity.

Reagents that are compatible with the RNaseAlert® technology are the ones marked "-" in the "Before Addition of RNase A" column and "+" in the "After Addition of RNase A" column.

This is not a comprehensive list of compatible reagents. It serves to illustrate some typical data using the RNaseAlert® Technology. More sophisticated monitoring can be performed with the RNaseAlert® QC System or RNaseAlert® QC System v2 using a fluorometer. Note that tips and solid surfaces are frequently positive for RNases if gloveless hands have simply touched them.

Ambion Kit Components Tested with the RNaseAlert® Technology Before Addition of RNase A* After Addition of RNase A* Comments
DNaseZap™ Solution 1 - +  
Gel Loading Buffer II - - Not recommended. The dye in the loading buffer quenches RNaseAlert® fluorescence.
MAXIscript™ Transcription Buffer, 1X - +  
MEGAscript™ Reaction Buffer, 1X - +  
NorthernMax™ Transfer Buffer - +  
NorthernMax™-Gly Gel Loading Buffer - - Inhibits RNase A activity.
NorthernMax™-Gly Gel Running Buffer, 1X - +  
Random Decamers, 10X - +  
Salmon Sperm DNA (sheared) - +  
THE RNA Storage Solution - +  


*Reagents that are compatible with RNaseAlert® technology are the ones marked "-" in the "Before Addition of RNase A" column and "+" in the "After Addition of RNase A" column.

Chemicals and Reagents Tested with the RNaseAlert® Technology Before Addition of RNase A* After Addition of RNase A* Comments
Acetone 10% - +  
Acrylamide/Bis-acrylamide 19:1, 4% - +  
Acrylic acid - - Inhibits RNase A activity.
Agarose Gel Loading Buffer, 10% - +  
Ammonium acetate, 500 mM - - Inhibits RNase A activity.
Ammonium persulfate, 1% - - Inhibits RNase A activity.
ATP, 1 mM - +  
Betaine, 500 mM - +  
Bromophenol Blue, 0.08% - - Not recommended. Bromophenol blue quenches RNaseAlert® fluorescence.
Cesium Chloride - - Inhibits RNase A activity.
Chloroform - -  
Coommassie Destain - - Inhibits RNase A activity.
DEPC-treated water - +  
DMSO 10% - +  
EDTA 100-500 mM - + 500 mM inhibits RNase A activity.
Ethanol, 10% - +  
Ethidium bromide - + Not recommended. Ethidium bromide has orange/red fluorescence that makes reading the fluorescence of the RNaseAlert® substrate difficult.
Glycerol - +  
Glycerol, 10% - +  
Glycine - +  
GTP, 7.5 mM - +  
Guanidinium hydrochloride, 100-400 mM - + 400 mM inhibits RNase A activity.
Guanidinium thiocyanate, 100-400 mM - + 200 mM inhibits 5 pg of RNase A
Magnesium acetate, 200 mM - - Inhibits RNase A activity.
Magnesium acetate, 30-100 mM - +  
Magnesium chloride, 100 mM - +  
Mineral Oil - +  
MOPS Gel Running Buffer, 10X - +  
NorthernMax Buffer (10X) - - Inhibits RNase A activity.
NP40, 1% - +  
Nuclease-free water (not DEPC-treated) - +  
PBS, 0.1X - +  
PBS, 1X - +  
PEG 8000, 4% - +  
Phenol - - Incubation with RNase A yielded slight violet fluoresecence.
Potassium acetate, 100 mM, pH 8.0 - +  
Potassium chloride, 200 mM - - Inhibits RNase A activity.
Potassium phosphate, 100 mM - - Inhibits RNase A activity.
RNAlater - -  
SDS, 1% - +  
Sodium Acetate, 300 mM, pH 5.2 - - Inhibits RNase A activity.
Sodium Chloride, 10-200 mM - +  
Sodium Chloride, 500 mM - - Inhibits RNase A activity.
Sodium Deoxycholate, 0.4% - +  
Sodium phosphate, 2 mM - +  
TBE, 1X - +  
TE, pH 8 - +  
Tris, 100 mM, pH 8.0 - +  
Tris, 100 mM, pH 7.0 - +  
Tween, 1.0% - +  
Urea polyacrylamide gel solution, 6% - +  
Yeast Lysis Buffer - +  
Zinc acetate, 100 mM - - Inhibits RNase A activity.


*Reagents that are compatible with the RNaseAlert® technology are the ones marked "-" in the "Before Addition of RNase A" column and "+" in the "After Addition of RNase A" column.

Materials Tested with the RNaseAlert® Technology Before Addition of RNase A* After Addition of RNase A* Comments
1 week old coffee with mold + ND* Glowed like the moon.
Agilent Bioanalyzer electrodes, touched with bare hands + ND Positive.
Frank's bare hands ** + ND No RNase activity detected.
Gary's bare hands** + ND Slight glow.
George's bare hands + ND Glowed brightly.
LB with bacterial funk, contaminated + ND Glowed like a Texas firefly in August.
Pipet tips touched with bare hands + ND Positive.
Saliva + ND Positive.


* ND = Not determined.
** 100 µl of RNase-free water was placed in the palm of the subject's bare hand. 5 µl of that water was used in the assay according to the kit protocol.

Enzymes* Tested with the RNaseAlert®  Technology Before Addition of RNase A* After Addition of RNase A* Comments
BamH1, 100 U - +++  
DNase I, 10 U - +++  
E. coli S30 extract, 1/10th Volume + ND**  
Hind III, 100 U - +++  
Klenow Fragment (Exo-) (50 U) - +++  
Kpn I - +++  
Msp I - +++  
Mung Bean Nuclease + ND  
Nco I - +++  
Nde I - +++  
Nuclease-free water - +++  
Proteinase K - +++  
RNase A +++ ND  
RNase I +++ ND  
RNase T1 +++ ND  
S1 nuclease +++ ND  
Sac I - +++  
Sal I - +++  
Shrimp Alkaline Phosphatase - +++  
SuperTaq - +++  
T3 RNA Polymerase / placental ribonuclease inhibitor protein mix, 100 U / 30 U - +++  
T4 Polynucleotide Kinase - +++  
T4 RNA Ligase - +++  
T7 RNA Polymerase / placental ribonuclease inhibitor protein mix, 100 U / 30 U - +++  
Xba I - +++  


*Tested at 10%
** ND = Not determined.