| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
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. |
Glycerol Content |
Contains Glycerol |
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.
Store this enzyme at -20°C and avoid repeated freeze-thaw cycles to preserve catalytic activity. The product is shipped Dry Ice and remains stable for up to one year from the date of manufacture when stored under recommended conditions. Aliquoting the stock solution into single-use volumes is recommended for enzymes used infrequently to minimize thermal cycling of the bulk stock.
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. Always verify compatibility with your specific template, buffer, and downstream workflow.
One unit (U) is defined as the amount of enzyme that catalyzes the ligation of greater than 95% of HindIII-linearized lambda DNA fragments into high-molecular-weight concatemers in 30 min at 16°C in a 20 µL reaction volume.
This enzyme is produced as Recombinant (E. coli) and supplied as a Research Use Only (RUO) reagent. Each lot is subjected to activity assay, purity assessment by SDS-PAGE, and functional validation prior to release. A Certificate of Analysis (CoA) and Safety Data Sheet (SDS) are available on request.
Standard T4 DNA ligase requires overnight ligation at 16°C for optimal efficiency with cohesive ends. Quick Ligase formulations contain optimized buffer systems and enhanced enzyme concentrations that achieve >90% ligation efficiency within 5–15 minutes at room temperature, significantly accelerating cloning workflows without compromising junction fidelity.
Yeasen Biotechnology supports custom enzyme solutions across multiple service lines — from GMP-grade bulk supply to directed enzyme engineering. Contact BioHippo to discuss requirements and initiate a project inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen enzymes are available in GMP grade, manufactured in an ISO 13485-certified UCF.ME™ ultra-clean molecular enzyme facility with FDA Drug Master File (DMF) support.
- GMP-grade release testing and CoA documentation
- ISO 13485-certified production facility
- Scalable from milligram to multi-gram quantities
- Consistent lot-to-lot activity specifications
▶ Glycerol-Free & Custom Formulation
Glycerol-free enzyme formats are available for applications requiring lyophilization compatibility, liquid handling automation, or direct IVD master mix integration.
- Glycerol-free liquid format (standard and custom buffers)
- Lyophilization-ready enzyme preparation
- Custom reaction buffer optimization for specific assay conditions
- Compatible with freeze-drying workflows for point-of-care formats
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting (RDC) team delivers end-to-end solutions for molecular diagnostic product development, covering enzyme selection through clinical validation support.
- Enzyme selection and performance matching
- Primer/probe design and reaction buffer optimization
- Sensitivity, specificity, and precision validation studies
- Stability studies and SNP evaluation
- Instrument platform compatibility assessment
▶ ZymeEditor™ Enzyme Engineering
Yeasen's proprietary ZymeEditor™ directed evolution and rational design platform enables the development of custom enzyme variants with tailored performance characteristics not available in off-the-shelf products.
- Directed evolution for enhanced thermostability, processivity, or fidelity
- Rational design for altered substrate specificity or cofactor requirements
- Library screening from Yeasen's proprietary enzyme variant collection
- Scale-up to commercial quantities upon candidate confirmation
ⓘ Customization services are fulfilled by Yeasen Biotechnology. Lead times and minimum order quantities vary by service type. Contact BioHippo for project scoping and pricing.
