Product Class: Other

Thermolabile Exonuclease I
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Item# Description List Price Web Price Qty
M0568L Thermolabile Exonuclease I - 15,000 units $426.00
$383.40
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M0568S Thermolabile Exonuclease I - 3,000 units $106.00
$95.40
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

    Product Introduction

    Thermolabile Exonuclease I catalyzes the removal of nucleotides from single-stranded DNA (ssDNA) in the 3′ to 5′ direction at 37°C, and can be easily heat-inactivated at 80°C for 1 minute. This enzyme has no activity on linear/circular dsDNA or RNA. 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 single-stranded DNA from dsDNA

    Product Information

    Description

    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.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    NEBuffer™ 3.1 -20 10X
    Product Categories:
    Nucleases Products,
    DNA Modifying Enzymes

    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
    • Removal of single-stranded DNA from dsDNA

    Properties & Usage

      Unit Definition

      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™ 3.1
      Incubate at 37°C

      1X NEBuffer™ 3.1
      100 mM NaCl
      50 mM Tris-HCl
      10 mM MgCl2
      100 µg/ml BSA
      (pH 7.9 @ 25°C)

      Storage Temperature

      -20°C

      Storage Conditions

      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 min

      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.

      Product Notes

      1. This enzyme can be heat inactivated at 80°C for 1 minute, 65°C for 5 minutes or 60°C for 15 minutes.
      2. Thermolabile ExoI is active in the presence of 25 mM to 200 mM NaCl.
      3. If the amount of ssDNA to be degraded needs to be increased, we recommend adding more enzyme instead of extending the reaction time.

      Protocols, Manuals & Usage

      Protocols

      1. Protocol for removing ssDNA from dsDNA or RNA Samples (M0568)

      Tools & Resources

      Selection Charts

      • Common Applications for Exonucleases and Endonucleases
      • Properties of Exonucleases and Endonucleases

      Faqs & Troubleshooting

      FAQs

      1. How can I remove the remaining primers and dNTPs from a PCR reaction prior to DNA sequencing?
      2. Can Thermolabile Exonuclease I remove primers in nested PCR reactions?
      3. Will Thermolabile Exonuclease I degrade RNA?
      4. Can Thermolabile Exonuclease I be used to blunt dsDNA?
      5. Can Thermolabile Exonuclease I be heat inactivated?
      6. Can Thermolabile Exonuclease I be used with a double-stranded exonuclease to clean up plasmid preparations?
      7. Will Thermolabile Exonuclease I work in other buffers?

      Troubleshooting

      Quality & Safety

      Quality Assurance Statement

      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

      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]

      Certificate Of Analysis

      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]

      Safety DataSheets

      The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

      Legal and Disclaimers

      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

      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.