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Which Water to Use?

by Sapna Chacko


High quality, DEPC treated, and non-DEPC treated water

Certified free of endonuclease, exonuclease, and RNase activity

Available in sizes to suit all your experimental needs

RNase contamination in reagents used for RNA isolation and analysis can contribute to experimental inconsistency and sometimes even experimental failure. In trying to pinpoint the source(s) of RNase contamination, it is easy to overlook the water used in the experiment, either to prepare reagents, or to resuspend precipitated RNA. Even purified water can have a high pH and minerals that can interfere with reactions that require specific salt and pH conditions. Ambion offers three different grades of high quality water: DEPC-treated water, Nuclease-free water (not DEPC-treated) and RT-PCR grade water.


Ambion Certified and Guaranteed Nuclease-free
All Ambion water products are Type II 18 ohm filtered, and are certified to be nuclease-free. Our water undergoes state-of-the-art reverse osmosis filtration and deionization, followed by autoclaving and sterile filtration. Ambion's water purification equipment is subjected to routine maintenance and testing procedures according to rigid ISO 9001 specifications. Finally, in order to be certified nuclease-free, each lot of water is teste d for nuclease activity by incubating a sample overnight with a radiolabeled RNA or DNA probe. After incubation, the probe is run on a gel to detect any degradation from nuclease.

1. DEPC-treated Water - DEPC destroys enzymatic activity by modifying -NH2, -SH and -OH groups in RNases and other proteins. DEPC treatment is a very effective way to treat solutions that will contact RNA. Ambion's DEPC-treated water is autoclaved both before and after packaging to ensure sterility and complete inactivation of DEPC.

2. Nuclease-free Water (not DEPC-treated) - Ambion also offers nuclease-free water that is not DEPC-treated. Ambion's Nuclease-free Water is ideal for applications that are reported to be acutely sensitive to residual DEPC e.g., oocyte injection and other in vivo translation applications.

3. RT-PCR Grade Water - Ambion's RT-PCR grade water is tested for prokaryotic as well as eukaryotic genomic DNA contamination using a sensitive end-point PCR assay based on detection of rRNA genes. Prokaryotic DNA contamination is assessed by performing PCR with 16S rRNA primers, and eukaryotic DNA contamination is assessed by performing PCR with 18S rRNA primers. Each lot is also tested for nuclease contamination. This product is ideal for use in any PCR or RT-PCR application.


Bulk and Custom Orders
All of Ambion's water products can be packaged into larger custom sizes to fit your needs. Contact us at custom@ambion.com for more information.

Inactivate RNases in Solution

DEPC treatment of solutions that come in contact with RNA can be time-consuming, possibly hazardous, and only eliminates RNases present at the time of treatment. Some solutions (e.g., primary amine containing compounds such as Tris) cannot be treated with DEPC at all.

The RNAsecure Reagent eliminates these problems. RNAsecure is a unique nonenzymatic reagent that will irreversibly inactivate RNases in solution. Unlike DEPC, RNAsecure can be used with virtually any solution and does not require post-treatment autoclaving. The inactivation process can be repeated at any time to protect against introduced contaminants. RNAsecure does not interfere with downstream procedures, such as RT-PCR and in vitro transcription, and is also compatible with Tris and other solutions that cannot be treated with DEPC.

The RNAsecure Resuspension Solution contains the same active ingredients as the RNAsecure Reagent, but is supplied at working concentration for direct resuspension of RNA pellets.


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Ordering Information
Cat# Product Name Size 1964 RNaseAlert Lab Test Kit 25 rxns 1966 RNaseAlert QC System 5 x 96 assays 1970 DNaseAlert QC System 5 x 96 assays 7005 RNAsecure Reagent 1 ml 7006 RNAsecure Reagent 10 ml 7010 RNAsecure Resuspension Solution 10 x 1 ml 9916 DEPC-treated Water 5 x 100 ml 9920 DEPC-treated Water 500 ml 9922 DEPC-treated Water 1 L 9924 DEPC-treated Water 4 x 1 L 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 9935 RT-PCR Grade Water 10 x 1.75 ml 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
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