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| Product Name | Plant Tissue Direct PCR Kit (With Dye) |
| Catalog No. | RESK-0059 |
| Description | The Plant Tissue Direct PCR Kit (With Dye) allows direct PCR amplification from a wide range of plant leaf samples with broad adaptability and strong stability. Its unique lysis buffer system rapidly lyses diverse plant tissues and releases genomic DNA; there is no need to remove proteins, RNA, or secondary metabolites before using the released genomic DNA directly as a PCR template. Sample requirements are minimal, as little as 1 mm of plant leaf is sufficient. The supplied 2x Plant Master Mix offers excellent amplification compatibility and performs efficient, specific amplification directly on sample lysates. This 2x concentrated master mix contains all components needed for PCR except template and primers, greatly simplifying the workflow and reducing contamination risk. An integrated tracking dye allows PCR products to be loaded directly for electrophoresis. The kit is well suited for transgenic plant identification, plant genotyping, and related applications. |
| Intended Use | For direct PCR amplification from plant leaf tissues without DNA extraction or purification. Suitable for identification of transgenic plants, plant genotyping, and gene identification. |
| Principle / Technology | A plant-specific lysis buffer (Buffer P1) rapidly disrupts leaf tissue and releases genomic DNA; a neutralization buffer (Buffer P2) neutralizes the lysate to protect DNA and extend sample storage. The hot-start 2x Plant Master Mix, containing an electrophoresis tracking dye, tolerates residual plant impurities and amplifies target DNA directly from neutralized lysates. |
| Sample Type | Leaves from a wide range of plant species; as little as 1 mm of plant leaf tissue is required per test. |
| Performance Range / Specifications | Amplifies fragments up to 1 kb. Extension time 60 sec/kb. Annealing temperature Tm-(2-5) C. Compatible with 40-65% GC content. Hot-start enzyme; blunt-end products; includes electrophoresis tracking dye. Neutralized lysate should have a pH of approximately 7-8 (neutralize lysate with Buffer P2 strictly at a 5:1 ratio). Template input should be optimized within 5% of the reaction volume; 35-40 cycles are recommended. |
| Specificity | Verified across multiple plant species: direct amplification from leaf tissue lysates produced specific target bands consistent with those obtained from the corresponding purified plant genomic DNA templates (30 cycles). |
| Reaction Conditions / Protocol | Lyse approximately 1 mm of leaf tissue in Buffer P1, then neutralize the lysate with Buffer P2 strictly at a 5:1 (lysate:P2) ratio so the final pH is about 7-8. Use the neutralized lysate (no more than 5% of the reaction volume) as template with the 2x Plant Master Mix. Anneal at Tm-(2-5) C with an extension time of 60 sec/kb; 35-40 cycles are recommended (generally 5-10 more cycles than with purified DNA templates). |
| Components / Formulation | Buffer P1 Buffer P2 2x Plant Master Mix |
| Storage Conditions | Buffer P1: 2-8 C. Buffer P2: 2-8 C. 2x Plant Master Mix: -25 to -15 C, avoid repeated freeze-thaw cycles; for frequent use, short-term storage at 2-8 C is acceptable. |
| Shelf Life | 1 year for each component and the kit under the recommended storage conditions. |
| Package Specifications | 50 T; 200 T; 5 T |
| Product Form | Kit |
| Quality Control | Kit performance is verified by direct amplification from leaf lysates of multiple plant species, benchmarked against purified plant genomic DNA. |
| Concentration | Master mix supplied as a 2x concentrate |
| pH Range / Optimal pH | Neutralized lysis mixture should be approximately pH 7-8 |
| Expiration Date / Stability | Valid for 1 year under the recommended storage conditions. Neutralized lysis mixtures can be stored at 2-8 C for up to 5 days; freshly prepared lysate is recommended for PCR. |
| Compatibility | Suitable for leaves of diverse plant species; the integrated tracking dye allows direct gel loading of PCR products. |
| Application Notes / Precautions | No bands in positive control and samples: optimize PCR conditions using a gradient cycler; store the 2x PCR mix at -25 to -15 C and avoid repeated freeze-thaw; redesign primers if necessary. Positive control amplifies but samples do not: use fresh reagents; neutralize lysate with Buffer P2 strictly at a 5:1 ratio (final pH about 7-8); use freshly prepared lysate (stable at 2-8 C for up to 5 days); optimize template input below 5% of the reaction volume; increase cycle number (35-40 cycles recommended, typically 5-10 more cycles than for purified DNA). Non-specific amplification: raise the annealing temperature; reduce cycle number, primer concentration, or template concentration; redesign primers; prepare reactions at low temperature and start PCR promptly after setup. Bands in the negative control: autoclave all reagents and consumables and handle gently to avoid aerosol contamination; use a separate sampling tool for each sample, or decontaminate blades by rinsing in 2% sodium hypochlorite and wiping dry between samples. |
For research use only, not for clinical use.
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