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| Product Name | Animal Tissue Direct PCR Kit (With Dye) |
| Catalog No. | RESK-0058 |
| Description | The Animal Tissue Direct PCR Kit (With Dye) enables direct PCR amplification from a wide variety of animal tissues, offering broad applicability, strong stability, and simple operation. A unique lysis buffer system rapidly lyses diverse animal tissue samples, such as insect legs and wings, mouse tails, mouse toes, and animal skin and viscera, releasing genomic DNA within minutes. The lysate can be used directly as a PCR template, subjected to further DNA extraction and purification, or stored at -20 C or below for later use. The supplied 2x Tissue Direct PCR Mix (With Dye) provides exceptional amplification compatibility: there is no need to remove tissue debris or impurities from the lysate, and efficient, specific amplification can be performed directly on the sample lysate. This 2x concentrated master mix contains all components required for PCR except template and primers, greatly simplifying the workflow and reducing the risk of contamination. The built-in tracking dye allows PCR products to be loaded directly onto gels. The kit is ideal for animal gene amplification and genotyping of transgenic animals. |
| Intended Use | For direct PCR amplification from animal tissues without DNA purification. Suitable for animal gene amplification analysis and genotyping of transgenic animals, including knockout mouse genotyping. Also applicable to direct PCR from animal cells (10^5 to 10^8 cells). |
| Principle / Technology | A dedicated lysis buffer system (lysis buffer plus proteases) rapidly digests animal tissue and releases genomic DNA. The inhibitor-tolerant 2x Tissue Direct PCR Mix (With Dye), a hot-start 2x master mix with an integrated electrophoresis tracking dye, amplifies target DNA directly from crude lysates without purification. A 5x PCR Enhancer is included for difficult templates. |
| Sample Type | Animal tissues such as insect legs and wings, mouse tail, mouse toe, animal skin, and visceral organs; also suitable for cultured animal cells at 10^5 to 10^8 cells per reaction. |
| Performance Range / Specifications | Amplifies fragments up to 1 kb. Extension time 20 sec/kb. Annealing temperature Tm-5 C. Compatible with 30-65% GC content. Hot-start enzyme; blunt-end products; includes electrophoresis tracking dye. Template preparation completed in 10 min; PCR finished in as little as 50 min. Lysis can be performed in 96-well plates for high-throughput workflows. |
| Specificity | Demonstrated specific amplification in genotyping of knockout mouse tissues: homozygous knockout (single 580 bp band), wild type (single 180 bp band), and heterozygous (580 bp/180 bp double bands) were clearly distinguished at 35 cycles, matching results obtained with 50 ng of purified genomic DNA. |
| Reaction Conditions / Protocol | Lyse tissue samples in the lysis buffer with proteases (no cutting or grinding required); template preparation is complete in about 10 min. Use the lysate directly as template with the 2x Tissue Direct PCR Mix (With Dye); add the 5x PCR Enhancer for challenging templates. Anneal at Tm-5 C with an extension time of 20 sec/kb; approximately 35 cycles are recommended for genotyping. Lysates may be stored at -20 C or below for long-term use. |
| Components / Formulation | Lysis Buffer Proteases 2x Tissue Direct PCR Mix (With Dye) 5x PCR Enhancer |
| Storage Conditions | Lysis Buffer: 2-8 C. Proteases: -25 to -15 C; do not leave the cap open for extended periods and avoid repeated freeze-thaw cycles. 2x Tissue Direct PCR Mix (With Dye): -25 to -15 C, avoid repeated freeze-thaw cycles. 5x PCR Enhancer: -25 to -15 C, avoid repeated freeze-thaw cycles. |
| Shelf Life | 1 year. |
| Package Specifications | 50 T; 200 T |
| Product Form | Kit |
| Quality Control | Kit performance is verified by direct amplification and genotyping of knockout animal tissue lysates, benchmarked against purified genomic DNA templates. |
| Concentration | PCR mix supplied as a 2x concentrate; enhancer supplied as a 5x concentrate |
| Expiration Date / Stability | Valid for 1 year under the recommended storage conditions. |
| Compatibility | Lysates can be used directly for PCR, further DNA extraction and purification, or long-term storage at -20 C or below. Compatible with 96-well plate lysis for high-throughput processing. |
| Application Notes / Precautions | This kit uses a protease-based (proteinase K-type) lysis method; an alkaline-lysis format is available in the dedicated mouse tissue direct PCR kit. Direct PCR from animal cells is supported; use 10^5 to 10^8 cells per reaction. Tissue samples do not need to be cut or ground before lysis, and the premixed PCR mix minimizes pipetting steps. |
For research use only, not for clinical use.
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