| Field | Specification |
|---|---|
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Quick T4 DNA Ligase is a single enzyme product that can be used for the ligation of DNA fragments and adapters in the process of NGS library construction. This product has been verified by high-throughput sequencing and has excellent quality. For customers who do not need experimental system adjustment, it is recommended to purchase Hieff NGSTM Fast-Pace DNA Ligation Module (Cat#12607).
Product Components
|
Component number |
Components |
10301ES40 |
10301ES42 |
|
10301-A |
Quick T4 DNA Ligase (400 U/µL) |
100 µL |
1 mL |
|
10301-B |
10 × T4 DNA Ligase Buffer |
250 µL |
2×1,250 µL |
Shipping and Storage
The product is shipped with ice packs and can be stored at -20°C for two years.
Unit Definition
In a 20 μL ligation reaction system, when 6 μg of λDNA-Hind Ⅲ is reacted at 16℃ for 30 mins, the amount of enzyme required to give more than 50% ligation of the DNA fragments was defined as one unit (U).
Cautions
1. If a small amount of precipitation occurs when the buffer is melting, please invert and mix well before use;
2. For your safety and health, please wear lab coats and disposable gloves for operation.
3. This product is for research use ONLY!
Instructions
1. The current mainstream DNA library construction kits are generally not purified after end repair and A-Tailing treatment, and directly perform adapter ligation. Please refer to the following preparation for the reaction system. Vortex thoroughly, spin briefly and incubate for 15 min at 20°C.
|
Components |
Volume (μL) |
|
dA-tailed DNA |
60 |
|
10 × T4 DNA Ligase Buffer |
10 |
|
50% PEG 6000 |
10* |
|
DNA Adapter |
X** |
|
Quick T4 DNA Ligase |
1-5*** |
|
ddH2O |
Up to 100 |
[Note]: *50% PEG 6000 is not provided in the kit, you need to prepare it by yourself.
**Refer to the following table for the amount of adapters.
***The amount of Quick T4 DNA Ligase used can be added 1-5 μL as needed.
2. The quality and concentration of adapters used directly affect ligation efficiency and library yield. Too much adapter usage may produce more adapter dimers, while lower usage may affect ligation efficiency and library yield. The following table lists the recommended amount of adapters used in different Input DNA situations using this kit.
|
Input DNA |
Adapter: Input DNA (molar ratio) |
Input DNA |
Adapter: Input DNA (molar ratio) |
|
1 μg |
10:1 |
50 ng |
100:1 |
|
500 ng |
20:1 |
25 ng |
200:1 |
|
250 ng |
40:1 |
1 ng |
200:1 |
|
100 ng |
100:1 |
500 pg |
400:1 |
[Note]: Input DNA mole (pmol)≈ Input DNA(ng)/ [0.66 × Input DNA average length(bp)].
Adapter ligation calculation example: How much adapter should be added when input DNA is 100 ng and length is 300 bp?
Step 1: Calculate the number of moles of input DNA. Formula: Input DNA mole (pmol)≈ Input DNA(ng)/ [0.66 × Input DNA average length(bp)]; Input DNA mole (pmol)=100 ÷ (0.66×300) = 0.5 pmol;
Step 2: According to the above table, calculate the number of moles of adapter added.; When the Input DNA is 100ng, the molar ratio of adapters: input DNA is 100:1, and the number of moles of adapters added=100×0.5 pmol=50 pmol;
Step 3: Calculate the adapter addition volume. Adapter concentration = 15 μmol/L (if you use other company's adapter, you need to determine its concentration according to its instructions); Volume of adapter added = moles of adapter added (50 pmol) ÷ concentration of adapter (15 μmol/L) = 3.34 μL (Note: 15 μmol/L = 15 pmol/ μL);
In summary, the volume to which the adapter can be added is 3.4 μL. (Note: The maximum addition volume of the adapter does not exceed 5 μL).
ase buffer.
Glycerol Content: Contains Glycerol
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.
This enzyme is validated for use in Cloning & Assembly, NGS Library Preparation applications. It is suitable for use in both standard laboratory research settings and, where applicable, optimized reaction workflows requiring high specificity, sensitivity, or throughput.
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.
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