Glycerol-Free T4 DNA Ligase (HC)

Glycerol-Free T4 DNA Ligase (HC) is a glycerol-free, high concentration enzyme (50U/µL) that catalyzes the formation of a phosphodiester bond between adjacent 5ʹ phosphate and 3' hydroxyl groups in duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

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Meridian's Glycerol-Free T4 DNA Ligase (HC)

  • Efficiently joins blunt and cohesive ends.

  • Repairs single-stranded nicks in duplex DNA, RNA and DNA/RNA hybrids.

  • Joins double-stranded oligonucleotide linkers or adaptors to DNA, for use in NGS library construction and cloning.

  • Glycerol-free, for lyophilization and ambient temperature storage, ideal for point of care (POC) assays.

Lyophilized Glycerol-free T4 DNA Ligase (HC) maintains its activity

Lyophilized Glycerol-free T4 DNA Ligase (HC) maintains its activity graph
Stability of Glycerol-free T4 DNA Ligase (HC) at room temperature in liquid or lyophilized format. For each condition, 330 units of T4 DNA Ligase was assessed for its ability to ligate 750 ng HindIII digested λ DNA in a standard ligation reaction for 1 hour at RT which was subsequently run on an agarose gel. ImageJ analysis was used to assess ligation efficiency relative to T4 DNA Ligase (T0). The results demonstrate that Glycerol-free T4 DNA Ligase (HC) is not affected by lyophilization and retains its activity when rehydrated.

Glycerol-free T4 DNA Ligase (HC), MDX200

Available in 1,000 Unit or 10,000 Unit aliquots

Lyo-Ready T4 Ligase Buffer, 5x, MDX204

Available in 1 mL or 10 mL aliquots

Glycerol-free T4 DNA Ligase (HC) requires a ligase reaction buffer. The concentration of Glycerol-free T4 DNA Ligase (HC) (in Weiss units) is higher than the concentration of other ligases on the market and can be lyophilized, making it ideal for microfluidics and point of care testing.

FAQs: Glycerol-free T4 DNA Ligase (HC)

T4 DNA Ligase should be used to ligate cohesive ends (10 minutes at room temperature) or blunt ends (2 hours at room temperature) or if the ligation is to be done overnight. T4 DNA Ligase should be used if heat inactivation is required. Concentrated T4 DNA Ligase is suggested for ligating single base overhangs.

One Weiss unit is defined as the amount of enzyme required to convert 1 nmol of 32P-labeled inorganic pyrophosphate into Norit adsorbable material (activated carbon) in 20 minutes at 37°C, using specified reaction conditions (Weiss, B., et al., J. Biol. Chem., 243, 4543 (1968)).

However, as the Weiss unit is not a direct measurement of DNA ligation, but an indirect measurement of the enzyme adenylation reaction, followed by determining ATP-PPi exchange, Cohesive End Unit is also used, as this is the direct measurement of DNA ligation (a Cohesive End Unit deing defined as the amount of enzyme required to give 50% ligation of Hind III fragments of lambda DNA in 20 µL of 1X T4 DNA Ligase Buffer in 30 minutes at 16°C). One Cohesive End Unit equals 0.015 Weiss units, or one Weiss unit equals 67 Cohesive End Units.

Yes, heat at 65 °C for 10 minutes.

The general formulation of a 1x ligase reaction buffer is 50 mM Tris-HCl (pH 7.8), 10 mM MgCl2, 10 mM DTT and 10 mM ATP.
Both Mg and ATP are an essential cofactor for the reaction, two ATP molecules are consumed for each phosphodiester bond formed.

The increased concentration of the T4 DNA ligase is helpful in lower volume reactions where reagent volumes are limited, such as in microfluidic and cassettes in point of care devices.

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