| Field | Specification |
|---|---|
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Exonuclease I, sourced from a genetically engineered E. coli strain expressing the Exo I gene, exhibits exonuclease activity that digests single-stranded DNA from the 3' to 5' end. It sequentially releases deoxyribonucleotide 5'-monophosphates, preserving the integrity of the 5'-terminal dinucleotides. This enzyme is predominantly utilized for the breakdown and removal of primers following PCR amplification. It remains inactive towards double-stranded DNA and DNA strands with 3' hydroxyl ends that are blocked by phosphorylation or acetylation.
Features
No residual exonuclease (double-stranded), endonuclease, or RNase
Inactivated by simply treating it at 80°C for 15 minutes
Applications
Remove single-stranded primers from the PCR reaction system before Sanger DNA sequencing or SNP analysis
Remove single-stranded primers from the nested PCR reaction system
Eliminate linear single-stranded DNA from the sample, leaving behind double-stranded DNA
Specifications
|
Source |
E. coli |
|
Molecular Weight |
55 KDa |
|
Concentration |
20 U/μL |
|
Unit Definition |
One unit is defined as the amount of enzyme required to catalyze the release of 10 nmol of acid-soluble nucleotides from single-stranded [3H]-DNA at a concentration of 0.17 mg/ml in a 50 μL reaction system containing 1X Exonuclease I reaction buffer at 37°C within 30 minutes |
Glycerol Content |
Contains Glycerol |
Components
|
Components No. |
Name |
14535ES80 |
14535ES90 |
|
14535-A |
Exonuclease I (20 U/μL) |
250 μL |
|
|
14535-B |
10×Exonuclease I Reaction Buffer |
1 mL |
1 mL |
Shipping and Storage
This product should be stored at -25 ~ -15℃ for 2 years.
Figures
Figure1. Digestion capability of Exonuclease I on single-stranded DNA
Note: In a 20 μL reaction system, a final concentration of 15 pmol/μL single-stranded DNA substrate was added. Different amounts (0.63, 1.25, 2.5, 5, 10, 20 U) of Yeasen's and the Supplier A's Exonuclease I were used and incubated at 37°C for 15 minutes, followed by heat inactivation at 80°C for 15 minutes. Polyacrylamide gel electrophoresis was employed to detect the degradation of single-stranded DNA. The results demonstrated that Yeasen's Exonuclease I has the same ability to digest single-stranded DNA as Supplier A.
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.
Remove single-stranded primers from the PCR reaction system before Sanger DNA sequencing or SNP analysis Remove single-stranded primers from the nested PCR reaction system Eliminate linear single-stranded DNA from the sample, leaving behind double-stranded DNA. 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.
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