Meridian High-performing Enzymes Guide 0525

Next Generation Sequencing (NGS)

Glycerol-Free T4 Polynucleotide Kinase (HC)

Used in NGS library preparation, either separately or as part of an End-Repair Mix, where phosphates the 5’-hydroxyl terminus of double stranded DNA for subsequent ligation of adaptors by T4 DNA Ligase.

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A/ Increase in fluorescence over time and B/ phosphorylation reaction rate of Glycerol-Free T4 Polynucleotide Kinase (HC) ( blue ) and NEB’s T4 Polynucleotide Kinase ( red ), used to demonstrate that the enzymatic activity of Glycerol-Free T4 Polynucleotide Kinase (HC) is comparable to glycerol-containing enzymes from other suppliers.

Glycerol-Free T4 DNA Ligase (HC)

Used to catalyze the formation of a phosphodiester bond between adjacent 5’ phosphate and 3’ hydroxyl groups in duplex DNA, efficiently joining blunt and cohesive ends and repairs single stranded nicks in duplex DNA. Used to join adaptors to DNA, for use in NGS library construction. NUCLEASE-FREE LIGATION: No detectable exo- or endonuclease activity, ensuring optimal ligation efficiency

Glycerol-Free T4 DNA Ligase (HC) was incubated of for 16 hours at 37°C with 1 μ g of HindIII-digested λ DNA (Lanes 2 and 3) and supercoiled pUC19 DNA (Lanes 5 and 6). The results show no detectable exo- and endonuclease activities that may affect the ligation efficiency of the enzyme.