DNA dA-Tailing Kit
DNA Mod
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- Application
- DNA Modification (DNA Mod)
- Purpose
- The DNA dA-Tailing Kit efficiently adds a non-template dAMP (dA) to the 3' end of a blunt-ended DNA fragment.
- Characteristics
- This incorporated 3'-dA prevents concatemer formation and prepares the DNA fragment for subsequent ligation of adaptors or cloning vectors that have complementary 3'-dT overhangs. The Klenow Fragment (3´→ 5´ Exo-) in this kit adds the 3'-dA via its DNA polymerase activity. This enzyme lacks 5'→3' exonuclease activity and has mutations that effectively abolish the inherent 3'→5' exonuclease activity, thus preventing degradation of this 3'-dA. This kit has been optimized to prepare compatible overhangs for the next step of DNA sample preparation for next generation sequencing. The Klenow Fragment (3´→ 5´ Exo-) enzyme is also available as a stand-alone enzyme (Cat. No. E038).
- Unit Definition
- One unit is defined as the amount of Klenow Fragment (3'→5' Exo-) that catalyzes the incorporation of 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C using poly(dA-dT):poly(dA-dT) as a template:primer.
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- Comment
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- DNA sample preparation
- dATailing of 1 5 μg of blunt ended DNA
- Restrictions
- For Research Use only
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- Buffer
- 25 mM Tris-HCl ( pH 7.5), 0.1 mM EDTA, 1 mM DTT, and 50 % (v/v) Glycerol.
- Storage
- -20 °C
- Storage Comment
- Store all components at -20°C in a non-frost-free freezer. All components are stable for 1 year from the date of shipping when stored and handled properly.
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