| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Tth DNA Polymerase is a heat-resistant DNA polymerase discovered from the thermophilic bacterium Thermus thermophilus HB8. In the presence of Mg²⁺, this enzyme exhibits 5′-3′ DNA polymerase activity and 5′-3′ exonuclease activity, but lacks 3′-5′ exonuclease activity, making it widely applicable for PCR amplification reactions. In the presence of Mn²⁺, the enzyme demonstrates strong reverse transcriptase activity at temperatures of 55-70°C, making it suitable for one-step RT-PCR reactions.
Specification
Unit Definition: One unit (U) of activity is defined as the amount of enzyme that incorporates 10 nmol of total nucleotides into an acid-insoluble product in 30 minutes at 74°C, using activated salmon sperm DNA as the template/primer.
Component
|
No. |
Name |
Size |
|
|
14607ES72 (250 U) |
14607ES80 (1250 U) |
||
|
14607-A |
Tth DNA Polymerase (5 U/μL) |
50 μL |
250 μL |
|
14607-B |
5×Tth RT-PCR Buffer |
1 mL |
4×1.25 mL |
|
14607-C |
10×Tth PCR Buffer (with Mg2+) |
500 μL |
2×1.25 mL |
|
14607-D |
50mM Mg(OAc)2 |
250 μL |
1.25 mL |
|
14607-E |
dNTP Mixture (10 mM each) |
150 μL |
750 μL |
Shipping and Storage
Shippingwith dry ice. Store at -20°C, with a shelf life of 2 years.
Precautions
1) If a small amount of precipitate appears in the buffer upon thawing, this is a normal phenomenon. Please invert and mix thoroughly before use.
2) For your safety and health, wear a lab coat and disposable gloves during operation.
3) This product is for research use only!
Instructions
Recommended PCR Reaction Procedure:
1. Preparation of the Reaction System
|
Name |
Volume (μL) |
|
10×Tth PCR Buffer (with Mg2+) |
5 |
|
Tth DNA Polymerase (5 U/μL) |
0.5 |
|
dNTP Mixture (10 mM each) |
1 |
|
Upstream Primer 10μM |
2 |
|
Downstream Primer 10μM |
2 |
|
Template DNA |
50 pg- 1μg |
|
ddH2O |
to 50 |
[Note]: The amounts of Tth DNA Polymerase, upstream and downstream primers, and dNTP Mixture can be adjusted according to specific experimental needs.
2. Cycle Setup
|
Steps |
Temp(ºC) |
Time |
Cycles |
|
Pre-denaturation |
94 |
30 sec-5 min |
1 |
|
Denaturation |
94 |
30 sec |
35
|
|
Annealing |
50-70 |
30 sec |
|
|
Extension |
72 |
60 sec/kb |
|
|
Final Extension |
72 |
10 min |
1 |
[Note]: Reaction conditions can be adjusted according to specific experimental requirements.
Recommended RT-PCR Reaction Procedure:
1. Preparation of the Reaction System
|
Name |
Volume (μL) |
|
5×Tth RT-PCR Buffer |
10 |
|
Tth DNA Polymerase (5 U/μL) |
1.5 |
|
50mM Mg(OAc)2 |
2.5 |
|
dNTP Mixture (10 mM each) |
1.5 |
|
Upstream Primer 10μM |
2 |
|
Downstream Primer 10μM |
2 |
|
Template RNA |
50 pg- 1μg |
|
ddH2O |
to 50 |
[Note]: The amounts of Tth DNA Polymerase, upstream and downstream primers, and dNTP Mixture can be adjusted according to specific experimental needs.
2. Cycle Setup
|
Steps |
Temp (ºC) |
Time |
Cycles |
|
Reverse transcription |
55-70 |
30 min |
1 |
|
Pre-denaturation |
94 |
30 sec-5 min |
1 |
|
Denaturation |
94 |
30 sec |
35
|
|
Annealing |
50-70 |
30 sec |
|
|
Extension |
72 |
60 sec/kb |
|
|
Final Extension |
72 |
10 min |
1 |
[Note]: Reaction conditions can be adjusted according to specific experimental requirements.
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 PCR & Amplification 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 incorporation of 10 nmol of deoxyribonucleotide into acid-insoluble material in 30 min under standard assay 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.
This enzyme is produced recombinantly in E. coli under stringent quality controls and validated on multiple substrate types. Key performance parameters include specific activity, substrate specificity, cofactor requirements, optimal pH/temperature range, and compatibility with downstream assays. Refer to the product datasheet for validated lot-specific activity data and recommended reaction conditions.
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.
