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Glycerol-Free DNA Pol I Klenow Fragment exo- (HC)

Glycerol-Free DNA Pol I Klenow Fragment exo- (HC) is a high concentration N-terminal truncation of E.coli DNA Polymerase I which retains polymerase activity, but has lost 5’-3’ exonuclease activity and has mutations (D355A, E357A) which eliminates the 3’-5’ exonuclease activity. It is designed for dA and dT tailing for adapter ligation in next generation sequencing library preparation.

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Meridian's Glycerol-Free DNA Pol I Klenow Fragment exo- (HC)

  • dA- and dT-tailing for next-generation sequencing (NGS) adapter ligation

  • Labeling by fill-in 5'-overhangs of dsDNA

  • DNA sequencing by the Sanger method

  • Isothermal DNA amplification SDA (Strand Displacement Amplification)

Glycerol-Free DNA Pol I Klenow Fragment exo- (HC), MDX218

Available in 200 µL (10,000 Units) or 1 mL (50,000 Units) aliquots

Glycerol-Free DNA Pol I Klenow Fragment exo- (HC) is as a glycerol-free, high concentration enzyme (50 U/L) 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 exo- (HC) is in a glycerol-free storage buffer and is supplied with a Lyo-Ready Klenow Reaction Buffer (5x) 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.

FAQs: Glycerol-Free DNA Pol I Klenow Fragment exo- (HC)

Just as the 5′ – 3′ exonuclease activity of DNA polymerase I from E.coli can be undesirable, the 3′ – 5′ exonuclease activity of Klenow fragment can also be undesirable for certain applications. This problem can be overcome by introducing mutations in the gene that encodes Klenow. This results in forms of the enzyme being expressed that retain 5′ – 3′ polymerase activity but lack any exonuclease activity (5′ – 3′ or 3′ – 5′).

No, DNA Pol I Klenow Fragment exo- cannot fill in 3′ overhangs. To create blunt ends 3′ overhangs must be removed. Glycerol-Free DNA Pol I Klenow Fragment (HC) (MDX208) and T4 DNA Polymerase (MDX207), are the best choices to remove 3′ overhangs.

Yes, DNA Pol I Klenow Fragment exo- is ideal for this application, it does so by filling in 5’ overhangs, and the lack of 3’→5′ exonuclease activity prevents the labeled nucleotides from being removed after they are incorporated by the polymerase.

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Have questions about a product? Want to learn more about Meridian’s molecular or immunoassay reagent portfolio? We want to hear from you!


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