Glycerol-Free DNA Pol I Klenow Fragment (HC)
Glycerol-Free DNA Pol I Klenow Fragment (HC) is an N-terminal truncation of E.coli DNA Polymerase I which retains polymerase and 3’➔ 5’ exonuclease activity but has lost 5’➔ 3’ exonuclease activity. It is designed for DNA blunting by 3’ overhang removal and fill-in of 5’ overhangs for adapter ligation.
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Glycerol-Free DNA Pol I Klenow Fragment (HC), MDX208
Available in 200 µL (10,000 Units) or 1 mL (50,000 Units) aliquots
Glycerol-Free DNA Pol I Klenow Fragment (HC) is a high concentration enzyme (50 U/µL) and is mainly used in next-generation sequencing (NGS) library preparation, where it can be used separately or as part of an End-Repair Mix, for double stranded DNA blunting by 3’ overhang removal and fill-in of 5’ overhangs for subsequent ligation of adaptors by T4 DNA Ligase. The enzyme displays moderate strand displacement activity and can also be used to generate probes using random primers, dideoxy sequencing and random priming labelling.
Glycerol-Free DNA Pol I Klenow Fragment (HC) is in a glycerol-free storage buffer and is supplied with a Lyo-Ready™ Klenow Reaction Buffer and has been optimized to deliver excellent, stable performance without glycerol, allowing you to deliver reliable results with the added capability for lyophilization. This gives you the flexibility to confidently store mixtures at ambient temperature or produce diagnostic assays that rely on miniaturized reaction components, such as point-of-care NGS.
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FAQs: Glycerol-Free DNA Pol I Klenow Fragment (HC)
No, DNA polymerases cannot remove 3′ overhangs, 3′ overhangs are removed by T4 DNA Polymerase (MDX207).
We recommend 25°C, higher temperatures can cause excessive 3’ ➔ 5’ exonuclease degradation, which eliminates blunt end formation.
Klenow Fragment is the large fragment of DNA Polymerase I that retains its 5’➔3′ polymerase, 3’➔5′ exonuclease and strand displacement activities. The enzyme lacks the powerful 5’➔3′ exonuclease activity of intact DNA polymerase I. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5′ termini.
Klenow Fragment can be used at 25°C or room temperature, but T4 DNA Polymerase must be used at 12°C due to its robust exonuclease activity.
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