| Field | Specification |
|---|---|
| Mfr No | |
| Form | SuperMix or Master Mix |
| Product Type | |
| Shipping | |
| Storage |
2× Hieff™ HotStart PCR Genotyping Master Mix is a ready-to-use 2× premix containing Hieff™ HotStart Taq DNA Polymerase, dNTPs, and an optimized buffer system. Only primers and template are needed to begin amplification, enabling high-throughput genotyping with improved reproducibility. The HotStart Taq polymerase is ligand-modified and completely blocked at room temperature; activity is released by a 2–3 minute 95°C activation step compatible with standard PCR programs.
Features
- Ready-to-use 2× premix — add only primers and template
- Hot Start technology prevents non-specific amplification during reaction assembly
- Rapid 2–3 min activation at 95°C; compatible with standard PCR protocols
- Thermostable with extended half-life at 95°C
- Generates PCR products with 3′-dA overhangs for TA cloning
Input DNA Amounts
| Sample Type | Recommended Input |
|---|---|
| Genomic DNA | 50–100 ng |
| Plasmids | 100 pg–20 ng |
| cDNA | 1–5 µL |
PCR Cycling Conditions
| Step | Temperature | Time | Cycles |
|---|---|---|---|
| Initial Denaturation | 95°C | 5 min | 1 |
| Denaturation | 95°C | 30 sec | 35 |
| Annealing | 50–60°C | 30 sec | 35 |
| Extension | 72°C | 30 sec/kb | 35 |
| Final Extension | 72°C | 10 min | 1 |
Selected Citations
[1] Han P, et al. Knockdown of hnRNPA1 promotes NSCLC metastasis and EMT by regulating alternative splicing of LAS1L exon 9. Front Oncol. 2022;12:837248. (IF: 5.738)
The Hieff™ HotStart Taq DNA Polymerase is ligand-modified, completely blocking polymerase activity at room temperature. Activity is fully released by heating at 95°C for 2–3 minutes. This prevents non-specific amplification and primer-dimer formation during reaction assembly, making it particularly effective for multiplex genotyping with multiple primers.
Store at −20°C. The formulation is stable under recommended storage conditions for at least one year from the date of manufacture. The premix format eliminates separate enzyme and buffer management.
Recommended inputs: genomic DNA 50–100 ng per reaction; plasmid DNA 100 pg–20 ng; cDNA 1–5 µL. Exceeding recommended inputs may reduce specificity.
This mix generates PCR products with 3′-dA overhangs, compatible with TA cloning and T-vector systems. The hot start mechanism does not affect product end chemistry.
Yes. The 2–3 minute 95°C activation step is fully compatible with existing standard PCR cycling programs. Simply insert an initial denaturation step of 95°C for 5 minutes at the start of your existing protocol.
Yeasen Biotechnology supports custom formulation and scale-up services for reagent mixes and master mixes. Contact BioHippo to discuss project requirements and initiate an inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen reagent mixes are available in GMP grade for IVD assay development. Manufactured in an ISO 13485-certified UCF.ME™ facility with FDA Drug Master File (DMF) support.
- GMP release testing and CoA documentation
- Scalable from milliliter to liter-scale quantities
- Consistent lot-to-lot performance specifications
▶ Glycerol-Free & Lyophilization-Ready Formats
Glycerol-free and lyophilization-ready formats are available for reagents intended for cartridge-based point-of-care devices, liquid handling automation, or ambient-temperature distribution.
- Glycerol-free liquid formulation
- Lyophilization-compatible format with cryoprotectant
- Custom reaction buffer optimization
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting team provides end-to-end support for molecular diagnostic product development using Yeasen reagent mixes as a foundation.
- Master mix selection and performance matching for target assay
- Primer/probe design and assay buffer optimization
- Sensitivity, specificity, and LOD validation
- Instrument compatibility assessment
▶ Custom Formulation
Custom formulation services are available for researchers requiring modifications to standard reagent mix compositions, including ROX level adjustment, dye substitution, stabilizer optimization, and buffer modification.
- ROX concentration customization (No / Low / High ROX)
- Reference dye and indicator dye options
- Buffer composition and pH optimization
- Scale-up to commercial quantities on confirmation
ⓘ Customization services fulfilled by Yeasen Biotechnology. Contact BioHippo for project scoping and pricing.
