Product Class: Kit

Acyclonucleotide Set

Product Introduction

Acyclonucleotide Set contains four separate tubes of acyNTPs (acyATP, acyCTP, acyGTP and acyTTP). 
Acyclonucleotides (acyNTPs) act as chain terminators and are thus useful in applications that normally employ dideoxynucleotides such as DNA sequencing (1,2) and SNP detection (3). AcyNTPs are especially useful in applications with archaeon DNA Polymerases, more specifically with Therminator™ DNA Polymerase. Therminator DNA Polymerase is an engineered enzyme with an increased capacity to incorporate analogs with altered sugars, such as ribonucleotides, dideoxynucleotides, 2´ deoxynucleotides and especially acyclo-base analogs (4,5). 
Catalog # Size Concentration
N0460S 0.5 µmol

Product Information

Description

When used with Therminator DNA Polymerase (NEB #M0261), the concentration of acyclo-base analog is roughly equivalent to dideoxynucleotide concentrations used with Thermo Sequenase™ (see figure).

(50 ng/µl single-stranded M13mp18 DNA, 0.05 µM [32P]-S1224S primer, 1X ThermoPol Buffer, 50 µM dNTP, varying acyGTP and 0.05 units/µl Therminator) were mixed, incubated at 94°C for 5 minutes and then thermal cycled for 25 cycles at 94°C (30 seconds); 55°C (30 seconds); 72°C (30 seconds). Reaction products were separated on a denaturing polyacrylamide gel (QuickPoint, NOVEX, Inc.) and analyzed by autoradiography. Numbers above each lane give the molar ratio of acyGTP to dNTP in each lane (i.e., 50, 17, 5,6 and 1.9 µM acyGTP, respectively). Numbers to the side of the gel indicate the length of the primer extension product added to the primer.
This product is related to the following categories:
Nucleotide Solutions Products
This product can be used in the following applications:
DNA Sequencing,
PCR,
PCR

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • N0460S     -20    
      AcycloATP N0461AVIAL -20 1 x 0.5 µmol Not Applicable
      AcycloCTP N0462AVIAL -20 1 x 0.5 µmol Not Applicable
      AcycloGTP N0463AVIAL -20 1 x 0.5 µmol Not Applicable
      AcycloTTP N0464AVIAL -20 1 x 0.5 µmol Not Applicable

Properties & Usage

Product Notes

  1. Thermo Sequenase™ is a trademark of GE Healthcare.
  2. Addition of 50 μl of distilled or deionized (Milli-Q™) water will result in a final concentration of 10 mM acyNTP. The hydrated acyNTP solution should subsequently be stored at -20°C.

References

  1. Sanger, F., Nicklen, S. and Coulson, A.R. (1977). Proc. Natl. Acad. Sci. USA. 74, 5463-5467.
  2. Trainor, G.L. (1996). U.S. Patent No. 5,558,991.
  3. Haff, L.A. and Simirnov, I.P. (1997). Genome Methods. 7, 378-388.
  4. Gardner, A.F. and Jack, W.E. (1999). Nucl. Acids Res.. 27, 2545-2553.
  5. Gardner, A.F. and Jack, W.E. (2002). Nucl. Acids Res.. 30(2), 605-613.

FAQs & Troubleshooting

FAQs

  1. Are substrates terminated with 3' acyclonucleosides resistant to 3'-5' exonuclease activity?
  2. Which DNA polymerases incorporate acyclonucleotides?
  3. Can acyclonucleotides be used in Sanger sequencing?
  4. Can acyclonucleotides be used for sequencing-by-synthesis?

Quality, Safety & Legal

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

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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