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Cat. No.: HFDL-2k (for 2KU)

Cat. No.: HFDL-10k (for 10KU)

Description

High-Fidelity DNA Ligase possesses ligase activity that utilizes ATP as a cofactor to form phosphodiester bonds by connecting the 5' phosphate end and 3' hydroxyl end of adjacent oligonucleotide chains that hybridize with the same complementary DNA strand. It requires both oligonucleotides to be fully paired with the complementary DNA strand, with no gaps between the two chains, for the reaction to occur. This enzyme exhibits excellent heat resistance and remains active within the temperature range of 45-70°C. Our company's high-fidelity DNA ligase is recombinantly expressed and prepared through multi-step purification of the recombinant protein.

Activity Definition

One unit is defined as the amount of enzyme required to ligate 50% of the λ DNA fragments (1 µg) digested with BstEII (approximately equivalent to 0.036 pmole of 12 bp sticky ends) within 15 minutes under the reaction conditions of 1× High-Fidelity DNA Ligase Buffer at a temperature of 45°C. Sticky end activity units are equivalent to cohesive end units.

Activity Assay Conditions

10 mM Tris-HCl, 600 µM ATP, 2.5 mM MgCl2, 2.5 mM DTT, 0.1% Triton X-100 (pH 7.5 @ 25°C), incubation at 45°C.

Concentration

40 U/μl

Features

• Excellent heat resistance, compatible with the template denaturation step in Ligase Chain Reaction (LCR).
• Extremely high fidelity, no mispaired base pairs at gapped positions will be ligated.
• Capable of ligating at cut sites on DNA strands under high-temperature conditions.
• Higher ligation specificity and lower non-specific background compared to room-temperature DNA ligases.
• Higher ligation specificity and lower non-specific background compared to Taq DNA ligase.
• Reaction temperature range of 35-70°C, with increased specificity and activity at higher temperatures, approaching 80°C.

Applications

• Detection of allele-specific genes through Ligase Detection Reaction (LDR) and Ligase Chain Reaction (LCR).
• Ligation of phosphorylated oligonucleotides in Ligase Chain Reaction (LCR).
• Ligation at cut sites on DNA strands under high-temperature conditions.

Storage

Store at -20°C.

DNA Ligase Products:

• T4 DNA Ligase
• Rapid T4 DNA Ligase
• T4 DNA Ligase (Fast)
• T7 DNA Ligase
• E. coli DNA Ligase
• PBCV-1 DNA Ligase
• T3 DNA Ligase
• Taq DNA Ligase
• Pfu DNA Ligase
• High-Fidelity DNA Ligase
• High-Salt DNA Ligase

SBS Genetech is recognized as one of the global major leading industry players in Ligase Market by third-party market researchers. For more details, please visit Ligase Market - A Global and Regional Analysis Focus on Product, Source, Application, End User, and Country - Analysis and Forecast, 2022-2032