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Product Overview
This kit is specially designed for mouse transgene identification and genotyping. It can directly and rapidly extract genomic DNA from mouse tissue (such as mouse tail, mouse ear, mouse toe, muscle, etc.) for subsequent PCR amplification and detection. The entire process does not require homogenization, disruption, overnight digestion, phenol-chloroform extraction, DNA precipitation or column purification. The sample requirement is small: 5–10 mg of mouse tissue or 1–5 mm of mouse tail is enough for genotyping.
The 2× Direct PCR Mix provided by this kit is a PCR master mix with high amplification compatibility, which can perform efficient and specific amplification without completely removing impurities such as proteins. The master mix contains antibody-modified Hot Star Taq DNA Polymerase, dNTPs, Mg²⁺ and other components, and only needs to add template and primers. The 2× Direct PCR Mix is pre-mixed with electrophoresis dyes, which can be directly detected by electrophoresis after the reaction is completed. The 3' end of the PCR product has an A, which can be used for TA cloning. Tissue is lysed at 55°C for 30 min and centrifuged at 12,000 rpm for 2 min before PCR; the recommended cycling is 94°C for 1–5 min, then 35 cycles of 94°C for 30 s, 60°C for 30 s and 72°C for 30 s/kb, with a final 72°C for 10 min.
Specifications
| Format | Direct PCR kit: Tissue Lysis Buffer, Digestive Enzyme, 2× Direct PCR Mix |
|---|---|
| Sample types | Mouse tissue such as tail, ear, toe and muscle |
| Sample input | 5–10 mg mouse tissue or 1–5 mm mouse tail |
| Polymerase | Antibody-modified Hot Star Taq DNA Polymerase (hot start) |
| Tracking dye | Electrophoresis dyes premixed |
| Reaction volume | 20 µL |
| Template input | 1–5% of the reaction volume |
| Amplicon length | Within 2 kb recommended |
| Product ends | 3' A overhang, suitable for TA cloning |
Kit Components
| Component | 50 T | 200 T |
|---|---|---|
| Tissue Lysis Buffer | 1.25 mL × 4 | 20 mL |
| Digestive Enzyme | 250 µL | 1 mL |
| 2× Direct PCR Mix | 500 µL | 1 mL × 2 |
| ddH₂O | 1 mL | 4 mL |
Applications
- Mouse transgene identification
- Mouse genotyping from tail, ear and toe tissue
- Direct PCR from mouse tissue without DNA purification
- TA cloning of PCR products
Key Features
- Simple and fast: suitable for one-step gene identification from mouse tail, mouse ear, mouse toe and other tissues.
- High specificity: This product uses antibody-modified hot-start Taq enzyme, which has high template affinity and amplification specificity.
Storage & Handling
All components are stored at -20°C for 3 years.
Usage Notes
- Avoid cross-contamination between samples.
- Use fresh tissue.
- Minimize freeze-thaw cycles.
- Establish negative and positive controls for each run.
- Keep the PCR product length within 2 kb for optimal efficiency.
Safety & Regulatory
For research use only. Not intended for clinical, diagnostic or therapeutic use. Follow the manufacturer's manual and standard laboratory safety practice.
Customization & Add-ons: Need this molecular biology reagent tailored to your workflow? We can help with bulk or custom pack sizes, alternative formats (spin column, magnetic bead or reagent-only kits; premixed or stand-alone reference dye), matched extraction, cDNA synthesis and qPCR workflows, and sourcing of related enzymes, ladders and consumables not in our catalog. Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services — our team will follow up with feasibility details and next steps.
