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TB #180: How to Maintain an RNase-free Lab Levels
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Technical Bulletin #180

How to Maintain an RNase-free Lab

Let's admit it, everyone who has worked with RNA has, at some point, been affected by RNase contamination. Sources of contaminating RNase include procedures that use RNase such as plasmid preps and nuclease protection assays. Bacterial cultures grown for plasmid preps or cloning, and bacterial products, such as some restriction enzymes can be a source of RNase. Microorganisms in the air, on surfaces, in one's solutions or water supply are environmental sources of RNase. Even you are a source of RNase. Human skin and body fluids such as saliva, tears and mucus, can contain RNases (so don't cry over your samples). Because RNases exhibit such a common presence in routine lab research, it is essential to develop a program of RNase control involving reagent preparation and testing, routine decontamination, and the use of certified RNase-free reagents and equipment such as microfuge tubes and pipette tips. See below for a schedule for maintaining an RNase-free lab. (For additional information on working with RNA, see Technical Bulletin #159: Working with RNA)

Schedule For RNase Control

Ambion scientists recommend the following schedule for RNase contamination control:

    Daily
  • Use Ambion's certified RNase-free buffers and reagents
  • Use Ambion's certified RNase-free consumables, such as microcentrifuge tubes and pipette tips
  • Use Ribonuclease Inhibitor Protein and/or ANTI-RNase in reactions such as in vitro transcription and RT-PCR
    Weekly
  • Thoroughly clean lab benchtops, pipettors (see sidebar below) and tube racks with RNaseZap®
    As Needed
  • Test bench-prepared reagents for RNase with RNaseAlert
  • Clean electrophoresis equipment with RNaseZap prior to use with RNA
  • Use aerosol resistant pipette tips when performing procedures that incorporate the use of RNase

How to Decontaminate Pipettors with RNaseZap®



Routine cleaning of pipettors with an RNase decontamination solution like Ambion's RNaseZap can greatly reduce the likelihood of sample loss due to RNase contamination. To decontaminate pipettors, carefully remove the pipettor shaft and ejector from the pipettor body. Be careful not to loosen the small seal and o-ring on the end of the plunger. Squirt RNaseZap into the shaft and wipe the shaft exterior and tip ejector with an RNaseZap-saturated paper towel. The same paper towel can be used to carefully wipe the pipettor body. Immediately wipe off any excess RNaseZap. Be careful that the paper towel is not so wet that liquid could seep into the pipettor. Rinse the shaft and ejector with RNase-free water to remove any RNaseZap residue. The water rinse can be followed by a brief ethanol rinse to expedite drying. Once shaft and ejector are dry, reassemble the pipettor.

 

Related Articles
Nuclease and Protease Testing

RNase and DEPC Treatment: Fact or Laboratory Myth?

Is Your RNA Intact?

RNase Control: The Basics

Top Ten Sources of RNase Contamination

Ordering Information
For prices and availability, please contact Customer Service.

Cat# Product Name Size
1964 RNaseAlert® Lab Test Kit 25 rxns
1966 RNaseAlert® QC System 5 x 96 assays
2682 RNase Inhibitor (Cloned) 40 U/µl 2500 U
2684 RNase Inhibitor (Cloned) 40 U/µl 10,000 U
2690 ANTI-RNase (15-30 U/µl) 2500 U
2692 ANTI-RNase (15-30 U/µl) 10,000 U
2694 SUPERase•In™ 2500 U
2696 SUPERase•In™ 10,000 U
7005 RNAsecure™ Reagent 1 ml
7006 RNAsecure™ Reagent 10 ml
7010 RNAsecure™ Resuspension Solution 10 x 1 ml
9010 Buffer Kit 100 ml ea
9780 RNaseZap® 250 ml
9782 RNaseZap® 6 x 250 ml
9784 RNaseZap® 4 L
9786 RNaseZap® Wipes 1 container (100 sheets)
9788 RNaseZap® Wipes Refill 3 x 100 sheets
9930 Nuclease-free Water (not DEPC-treated) 500 ml
9932 Nuclease-free Water (not DEPC-treated) 1 L
9934 Nuclease-free Water (not DEPC-treated) 4 x 1 L
9937 Nuclease-free Water (not DEPC-treated) 10 x 50 ml
9938 Nuclease-free Water (not DEPC-treated) 100 ml
9939 Nuclease-free Water (not DEPC-treated) 5 x 100 ml
12225 Thin-walled, Frosted Lid, RNase-free PCR Tubes 1000 x 0.2 ml
12250 Thin-walled, Dome Cap, RNase-free PCR Tubes 1000 x 0.5 ml
12275 Thin-walled, Frosted Lid, RNase-free PCR Tubes 1000 x 0.5 ml
12300 RNase-free Microfuge Tubes 1000 x 0.5 ml
12350 Non-Stick RNase-free Microfuge Tubes 500 x 0.5 ml
12400 RNase-free Microfuge Tubes 500 x 1.5 ml
12425 RNase-free Microfuge Tubes 500 x 2.0 ml
12450 Non-Stick RNase-free Microfuge Tubes 250 x 1.5 ml
12475 Non-Stick RNase-free Microfuge Tubes 250 x 2.0 ml
12480 Elution Tubes 100 x 2.0 ml
12635 Barrier (Filter) Tips, 10 µl size - Eppendorf Ten 8 x 12 racks
12640 Barrier (Filter) Tips, 10 µl size - Pipetman® Ten 8 x 12 racks
12645 Barrier (Filter) Tips, 20 µl size Ten 8 x 12 racks
12650 RNase-free Tips, 200 µl size Ten 8 x 12 racks
12655 Barrier (Filter) Tips, 200 µl size Ten 8 x 12 racks
12660 RNase-free Tips, 1000 µl size 10 x 100 ct racks
12665 Barrier (Filter) Tips, 1 ml size 10 x 100 ct racks
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