T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the 5'-phosphate and 3'-hydroxyl termini of adjacent nucleotides in double-stranded DNA or RNA with either cohesive or blunt ends. This reaction requires ATP as a cofactor. Additionally, T4 DNA Ligase can repair single-strand nicks in double-stranded DNA, double-stranded RNA, or DNA/RNA hybrids. Specifications Cat.No. 14966ES40 / 14966ES42 Size 40 KU / 400 KU Concentration 400 U/μL Source Recombinant E. coli strain carrying the T4 DNA ligase gene. Enzyme Activity Definition One unit is defined as the amount of DNA ligase required to achieve 50% ligation of 100 ng of cohesive-end DNA fragments in 50 µL of 1X DNA Ligase Reaction Buffer with incubation for 30 minutes at 23°C. Features Excellent Heat Stability: Maintains over 60% residual activity after 4 hours at 42°C. Ultra-Low Adapter Self-Ligation: Cleaner Libraries, Higher Sensitivity Excellent batch-to-batch consistency: Stable and uniform performance. Components Components No. Name 14966ES40 14966ES42 14966-A Premium T4 DNA Ligase (400 U/μL) 100 µL 1 mL 14966-B 2 × Rapid Ligation Buffer 1 mL 10 mL 14966-C 10 × T4 DNA Ligase Buffer 1 mL 10 mL [Note]: ① High-concentration Premium T4 DNA Ligase combined with 2× Rapid Ligation Buffer significantly enhances the speed and efficiency of blunt-end ligation. Therefore, prolonged incubation (>10 minutes) is not recommended, as it may substantially reduce the transformation efficiency of the ligation products. ② The 10× T4 DNA Ligase Buffer does not contain PEG and is compatible with standard ligation protocols that do not specify the use of rapid ligation buffer. Storage This product should be stored at -25~-15℃ for 2 years. Applications Routine molecular cloning High-throughput or automated library prep Low-input NGS applications cfDNA and liquid biopsy workflows Challenging ligation reactions requiring long incubation Protocols involving room-temperature operations Whole-genome library construction Figures 1. Exceptional Thermostability: Retains >60% Activity After 4 Hours at 42°C Figure 1. Thermostability assessment at 42°C and 45°C Yeasen Premium T4 DNA Ligase (Lots 1–3) and other suppliers were incubated at 42°C and 45°C for up to 4 hours. Activity was measured using a whole-genome library prep workflow with 1 µg bovine DNA. Yeasen ligase retained >60% activity at 42°C and outperformed competitors at 45°C, demonstrating superior thermostability. 2. Ultra-Low Adapter Dimerization: Minimal Self-Ligation with 0.5 ng gDNA in Library Prep Figure 2. Comparison of adapter dimerization rates Three batches of Yeasen Premium T4 DNA Ligase were compared with ligases from Supplier A and Supplier B using whole-genome library prep with 0.5 ng bovine DNA (Yeasen DNA Library Prep Kit, Cat# 12927ES). Yeasen ligase showed significantly lower adapter dimer formation, reducing non-specific ligation and improving usable sequencing yield. 3. Low DNA Residuals Figure 3. Host gDNA residual detection results for Premium T4 DNA Ligase. Three batches of Yeasen Premium T4 DNA Ligase were tested for E. coli host gDNA contamination using the Yeasen E. coli Host Cell DNA Residual Detection Kit (Cat# 41318). All batches exhibited extremely low residual gDNA, well below the levels of imported brands, demonstrating high enzyme purity and ensuring reliable experimental outcomes. 4. No Nuclease Contamination Figure 4. Nuclease Contamination Test of Premium T4 DNA Ligase. Three batches of Premium T4 DNA Ligase (6000 U each) were incubated with nucleic acid substrates and analyzed by agarose gel electrophoresis. No exonuclease or nicking activity was detected, confirming the absence of nuclease contamination and ensuring sample integrity. 5. High Ligation Efficiency Figure 5. Ligation-Mediated Circularization Efficiency Comparison. The circularization efficiency of Yeasen Premium T4 DNA Ligase (Cat# 14966) was compared with a conventional T4 DNA ligase(Cat#10301,Yeasen) using MGI libraries with either single-end or dual-index adapters. Circular products were pooled at equal mass and sequenced on the T7 sequencer. Yeasen 14966 consistently showed higher circularization efficiency and greater sequencing data yield than the conventional ligase. Notes 1. For your safety and health, wear a lab coat and disposable gloves while handling this product. 2. This product is intended for research use only. 3. It is normal for a small amount of precipitate to appear when the buffer is thawed. Simply invert and mix well before use. Documents: Safety Data Sheet 14966_MSDS_HB251029_EN.PDF Manuals 14966_Manual_Ver.EN20251029.pdf Related Blog Breakthrough in DNA Ligation: Premium T4 DNA Ligase Redefines Stability, Precision, and Ultra-Low Self-Ligation
