Interested in a 5-minute protocol? Try out the Exo-CIP™ Rapid PCR Cleanup Kit (NEB #E1050)
We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.
DNA specific exonuclease
Catalyzes the removal of nucleotides from linear single-stranded DNA in the 3' to 5' direction
Unlike Exonuclease I (NEB #M0293), Thermolabile Exonuclease I can be quickly heat inactivated at 80°C in 1 minute
Need help finding the right exonuclease for your experiments? Try Exo Selector
Thermolabile Exonuclease I is ideal for:
Removal of single-stranded primers in PCR reactions prior to DNA sequencing or SNP analysis
Removal of single-stranded primers for nested PCR reactions
Removal of linear single-stranded DNA, leaving behind double-stranded DNA in the sample
Thermolabile Exonuclease I catalyzes the removal of nucleotides from single-stranded DNA (ssDNA) in the 3′ to 5′ direction.
Figure 1: Heat inactivation profile of Thermolabile ExoI
Thermolabile ExoI can be heat inactivated quickly to speed up downstream applications. This data demonstrates that Thermolabile ExoI can be heat inactivated at 80°C for 1 min, 65°C for 4 min or 60°C for 15 min in both NEBuffer 3.1 as well as Standard Taq Reaction Buffer.
Product Source
An E. coli strain that carries a cloned Thermolabile Exonuclease I gene originating from a psychrophilic marine bacterium.
This product is related to the following categories:
One unit of Thermolabile Exonuclease I is defined as the amount of enzyme that will catalyze the release of 2 nmol of acid-soluble nucleotide in a total reaction volume of 100 μl in 6 minutes at 37°C in 50 mM Tris-HCl (pH 7.9), 100 mM NaCl, 10 mM MgCl2 and 100 μg/ml BSA with 0.17 mg/ml single-stranded [3H]-E. coli DNA.
Reaction Conditions
1X NEBuffer™ r3.1
Incubate at 37°C
1X NEBuffer™ r3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)
Storage Buffer
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
0.25 M NaCl
50% Glycerol
200 µg/ml BSA
pH 7.4 @ 25°C
Heat Inactivation
80°C for 1 minute
Unit Assay Conditions
50 mM Tris-HCl (pH 7.9), 10 mM MgCl2, 100 mM NaCl, 100 μg/ml BSA and 0.17 mg/ml single-stranded [3H]-E.coli DNA.
Advantages and Features
Application Features
Removal of single-stranded primers in PCR reactions prior to DNA sequencing or SNP analysis
Removal of single-stranded primers for nested PCR reactions
This enzyme can be heat inactivated at 80°C for 1 minute, 65°C for 5 minutes or 60°C for 15 minutes.
Thermolabile ExoI is active in the presence of 25 mM to 200 mM NaCl.
If the amount of ssDNA to be degraded needs to be increased, we recommend adding more enzyme instead of extending the reaction time.
Phosphorothioate (PT) bond is resistant to Thermolabile Exonuclease I cleavage. However, only one of the two chiral forms of the PT bond is resistant. To achieve complete protection against degradation at all PT bond positions, it is recommended to include five consecutive PT bonds in chemically synthesized DNA. Learn more here.
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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