| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
T5 Exonuclease is a double-stranded DNA-specific exonuclease and single-stranded DNA endonuclease. It can initiate digestion from the 5' end of single- or double-stranded DNA, as well as from gaps or nicks in linear or circular double-stranded DNA, but it does not degrade supercoiled double-stranded DNA. Therefore, T5 Exonuclease is widely used for removing incompletely ligated products from circularized dsDNA, degrading denatured DNA in alkaline extraction plasmids, degrading linear and nicked DNA in plasmid samples, and seamless cloning (Gibson assembly), etc.
Applications
In-Fusion Cloning
Degradation and alkaline lysis extraction of denatured DNA from plasmids
Specifications
|
Concentration |
10 U/µL |
|
Unit Definition |
One unit is defined as the amount of enzyme required to effect a change of 0.00032 A260 nm per minute at 37°C in the buffer |
Glycerol Content |
Contains Glycerol |
Components
|
Name |
14538ES80 |
14538ES90 |
|
|
14538-A |
T5 Exonuclease (10 U/µL) |
100 μL |
500 μL |
|
14538-B |
T5 Exonuclease Buffer(10×） |
1 mL |
1 mL |
Shipping and Storage
The products should be stored at -25℃ ~ -15℃ for 3 years.
Figures
Figure1. Cloning Situation of Different Fragment Connections
Note: In-Fusion Cloning was carried out using T5 Exonuclease, with the connection of 2, 3, and 4 fragments respectively. The experimental results demonstrate that the application of T5 Exonuclease in In-Fusion Cloning can achieve a 100% positive clone rate (12 out of 12)
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.
In-Fusion Cloning Degradation and alkaline lysis extraction of denatured DNA from plasmids. Always verify compatibility with your specific template, buffer, and downstream workflow.
One unit (U) is defined as the amount of enzyme that degrades 1 µg of DNA or RNA substrate to acid-soluble form in 30 min at 37°C under defined buffer conditions.
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.
Nuclease activity typically requires divalent metal ion cofactors (Mg²⁺ or Mn²⁺ at 1–5 mM). Activity is inhibited by chelating agents such as EDTA (≥1 mM), high salt concentrations, and reducing agents such as DTT at elevated concentrations. Heat inactivation (65–75°C for 15 min) is effective for most nucleases, allowing removal of enzyme activity after digestion without column purification.
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.
