Modifying Enzymes and Library Prep Enzymes
NGS library construction is mostly polymerase and ligase work — but the enzymes that get the DNA ready for that polymerase and ligase are where library quality is actually set. End repair, A-tailing, 5′ phosphorylation, adapter ligation, contamination control, RNA protection: each step has a specific enzyme, and substituting a lower-concentration or lower-purity version into that workflow shows up in library complexity and adapter ligation efficiency.
This collection covers seven modifying enzymes across the core NGS and molecular biology accessory workflow.
End repair and blunting T4 DNA Polymerase for 3′ overhang removal and 5′ overhang fill-in to generate blunt ends from restriction fragments, sheared DNA, or other irregular termini. Klenow Fragment (3′→5′ exo⁻) for fill-in without 3′ exonuclease activity — the standard choice for A-tailing after blunting.
5′ phosphorylation T4 Polynucleotide Kinase (T4 PNK) for phosphorylating 5′ hydroxyl termini on DNA or RNA — required for adapter ligation on dephosphorylated ends and for preparing substrates in ligation-based NGS workflows.
Adapter ligation T4 DNA Ligase (High Concentration) at 2,000,000 U/mL for efficient blunt-end and cohesive-end ligation, adapter ligation, and cloning of restriction fragments. High-concentration format reduces reaction volume requirements in library prep workflows.
Contamination control UDG (Uracil-DNA Glycosylase) for carry-over PCR contamination control — degrades uracil-containing PCR products from previous reactions before amplification begins.
RNA protection RNase Inhibitor (Mammalian) for protecting RNA during reverse transcription, in vitro transcription, and cell-free expression reactions where RNase contamination would degrade template.
Tagmentation and epigenomics Magic™ Tn5 Transposase — a naked, hyperactive Tn5 variant that inserts adapter sequences into nucleosome-free and linker DNA regions for ATAC-seq library construction and second-generation sequencing applications.
