Next-Generation Sequencing (NGS)

STEP 2 CREATE BLUNT ENDS

Glycerol-Free T4 DNA Polymerase (HC)

Cat #MDX207

VOLUME REACTIONS 50 μ L 500 Rxns 500 μ L 5,000 Rxns

Glycerol-Free T4 DNA Polymerase (HC) (MDX207) is supplied at concentration 20 U/ μ L and is a template dependent DNA polymerase that catalyzes the addition of nucleotides to the 3’ end of a DNA strand, extending it in the 5’ to 3’ direction. It also possesses 3’ to 5’ exonuclease activity, allowing it to remove nucleotides from the 3’ end of DNA strands but lacks a 5´to 3´ exonuclease activity. It can be used for generating blunt ends on any duplex DNA molecule and for labelling DNA by replacement synthesis. In NGS library preparation it can be used separately or as part of an End-Repair Mix, where it can blunt DNA ends of double-stranded DNA fragments for subsequent ligation of adaptors.

Product Highlights • Blunting of DNA ends by “filling in” or “polishing” the ends of double-stranded DNA fragments, for cloning or generating NGS libraries. • Sanger sequencing, to remove fluorescent dye-labeled nucleotides as they are incorporated into the growing DNA strand. • Concentration 20 U/ µ L

No loss of activity after lyophilization

(Right) Glycerol-Free T4 DNA polymerase (HC) (MDX207) was lyophilized in its reaction buffer, and the stability was tested in an accelerated stability study. The lyophilized MDX207 ( red ) was incubated at 37°C for 1 month and its DNA polymerization activity was tested against a non-lyophilized aliquot stored at -20°C ( blue ). Results suggest that the lyophilized MDX207 is active following accelerated stability tests with projected 12 months stability at ambient temperature.

Liquid | Lyophilized

Glycerol-Free DNA Pol I Klenow Fragment (HC)

Cat #MDX208

VOLUME REACTIONS 200 μ L 10,000 Rxns 1 mL 50,000 Rxns

Glycerol-Free DNA Pol I Klenow Fragment (HC) (MDX208) is supplied at 50 U/ μ L and is a large N-terminal truncated protein fragment produced from E. coli DNA polymerase I, which retains polymerase and 3’-5’ exonuclease activity but has lost 5’-3’ exonuclease activity. It can be used to fill in 5´-protruding ends and is ideal for DNA blunting by 3’ overhang removal and fill-in of 5’ overhangs prior to adapter ligation in next-generation sequencing library preparation.

Product Highlights • DNA blunting for NGS adapter ligation. • Labeling by fill-in of 5’-overhangs of dsDNA. • Concentration 50 U/ µ L

No loss of activity after lyophilization

Fast kinetics with high performance

Meridian | NEB | Thermo | Qiagen

(Above) Glycerol-Free DNA Pol I Klenow Fragment (HC) was used in a primer extension assay. Glycerol-Free DNA Pol I Klenow Fragment (HC) was used at 5 U final in reaction with 4 ng of template and Lyo-Ready Klenow Reaction Buffer and compared to other suppliers under the same conditions. The results demonstrate that Glycerol-Free DNA Pol I Klenow Fragment (HC) has a faster reaction rate and higher end fluorescence than most suppliers.

Liquid | Lyophilized

www.meridianbioscience.com/lifescience (Above) Accelerated stability studies of Glycerol-Free DNA Pol I Klenow Fragment (HC) at 37°C in liquid ( blue ) or lyophilized ( red ) format. The results demonstrate that Glycerol-Free DNA Pol I Klenow Fragment (HC) is active following accelerated stability tests with at least a projected 12 months stability at ambient temperature.

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