Hemo KlenTaq®

Description

Hemo KlenTaq is a truncated version of Taq DNA Polymerase, lacking the first 280 amino acids (1). Hemo KlenTaq also contains mutations that make it resistant to inhibitors present in whole blood. It can amplify directly from whole blood from humans and mice. Hemo KlenTaq tolerates up to 20% whole blood in a 25 µl reaction (30% in 50 µl reaction).

Amplification of human whole blood with Hemo KlenTaq. Percent blood present in sample and anticoagulant used are indicated in the legend. Ladder M is the 2-Log DNA Ladder (NEB #N3200).

Product Source

An E. coli strain that carries a mutant Taq DNA polymerase gene. The protein lacks the N-terminal 5´→ 3´ exonuclease domain and the gene has three internal point mutations.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
Hemo KlenTaq® Reaction Buffer Pack-205X

Advantages and Features

Features

  • Versatility and robustness of Taq DNA Polymerase to amplify DNA from whole blood.

Applications

  • Whole Blood PCR
  • Dried Blood Spot PCR
  • Primer extension

Properties and Usage

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.5% Tween® 20
0.5% IGEPAL® CA-630
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Resulting Ends

  • Single-base 3´ Overhangs

Unit Assay Conditions

1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

References

  1. Kermekchiev, M.B., et al. (2009). Nucleic Acids Res. 37, e40.
  2. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques . 15, 372-374.

Tech Tips

For best results, slowly pipette your blood sample into the reaction last and let it settle to the bottom of the reaction. The reaction should not be mixed once the blood is added.

Protocols

  1. PCR Protocol for Hemo KlenTaq®

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Endonuclease Activity (Nicking):
    The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
  • PCR Amplification (Whole Blood DNA):
    The polymerase is tested in a polymerase chain reaction (PCR) using whole blood containing inhibitors and specific primers, resulting in the expected product.
  • Single Stranded DNA Binding (FAM labeled oligo):
    The product is tested for its ability to produce a mobility shift when incubated using standard reaction conditions with a single stranded FAM labeled oligo.

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

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]

Safety Data Sheet

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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.