Mouse Tail Direct PCR Kit, IC-5561
Product Introduction
This product is a ready-to-use direct PCR kit specifically designed for rapid genotyping and genetic identification from mouse tail biopsies without the need for time-consuming DNA extraction or purification steps. The kit utilizes a proprietary lysis buffer system combined with a highly robust and inhibitor-tolerant hot-start DNA polymerase, enabling direct amplification of target genomic sequences from crude tail lysates. This streamlined workflow significantly reduces hands-on time and sample processing steps, allowing researchers to obtain genotyping results within 60-90 minutes from tissue sampling to gel analysis. The kit is optimized for routine transgenic and knockout mouse colony maintenance, eliminating the need for organic solvents, column-based purification, or overnight proteinase K digestions typically associated with traditional genomic DNA extraction protocols.
Product Features
1. Extraction-Free Workflow: Direct lysis of mouse tail biopsies eliminates DNA purification steps, reducing processing time to approximately 15-20 minutes for sample preparation.
2. Inhibitor-Tolerant Polymerase: Proprietary hot-start DNA polymerase formulation resists common PCR inhibitors present in crude tissue lysates, ensuring robust and reproducible amplification.
3. Rapid Genotyping: Complete workflow from tail sampling to PCR results achievable within 60-90 minutes, ideal for high-throughput colony screening.
4. High Sensitivity and Specificity: Reliable amplification of target sequences up to 2 kb from as little as 1-2 mm of tail tissue.
5. Cost-Effective: Significantly reduces reagent costs and labor compared to traditional extraction-based genotyping methods.
Specifications
Size: Available in 50 reactions, 200 reactions, and 500 reactions
Kit Components: Tissue Lysis Buffer, Protease Solution, 2X Direct PCR Master Mix (Hot-Start), Nuclease-Free Water
Storage and Stability
Storage Conditions: Store Tissue Lysis Buffer and Protease Solution at 4ˇăC. Store 2X Direct PCR Master Mix at -20ˇăC, protected from light. Nuclease-Free Water may be stored at room temperature or 4ˇăC.
Shelf Life: All components are stable for 12 months from the date of manufacture when stored as directed. Avoid repeated freeze-thaw cycles of the 2X Direct PCR Master Mix.
Protocol (For Reference Only)
Important: Use fresh or frozen tail biopsies. Minimize tissue size to 1-2 mm for optimal lysis. Avoid carryover of excessive hair or feces which may inhibit PCR.
1. Tissue Lysis: Place 1-2 mm mouse tail biopsy into a PCR tube containing 50-100 µL Tissue Lysis Buffer supplemented with 2-5 µL Protease Solution, mix briefly, and incubate at 55ˇăC for 10-15 minutes to release genomic DNA.
2. Protease Inactivation: Transfer the lysate to a 95ˇăC heat block or thermal cycler and incubate for 5 minutes to completely inactivate the Protease Solution, then centrifuge briefly to collect condensation and debris.
3. PCR Setup: In a clean PCR tube or plate, combine 1-2 µL of crude tail lysate supernatant with 10-25 µL of 2X Direct PCR Master Mix, forward and reverse primers (0.2-0.5 µM final each), and Nuclease-Free Water to the appropriate final reaction volume.
4. Thermal Cycling: Perform PCR amplification using standard cycling conditions optimized for specific primer pairs and amplicon length; typical cycling includes an initial denaturation at 94-95ˇăC for 3-5 minutes followed by 30-35 cycles of denaturation, annealing, and extension.
5. Analysis: Analyze PCR products by agarose gel electrophoresis with appropriate DNA size markers to confirm genotype based on expected amplicon sizes.
Precautions
1. Use clean instruments for tail biopsy collection to prevent cross-contamination between animals; change scalpel blades or scissors between samples.
2. Avoid overloading the PCR reaction with crude lysate; excessive lysate volume may introduce inhibitors and reduce amplification efficiency.
3. Store 2X Direct PCR Master Mix at -20ˇăC and avoid repeated freeze-thaw cycles; aliquot into smaller volumes for frequent use.
4. For difficult or GC-rich templates, optimization of annealing temperature or addition of PCR enhancers may be required.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the maximum amplicon size this kit can amplify?
A1: The kit reliably amplifies fragments up to 2 kb from crude tail lysates. For larger amplicons or difficult templates, optimization of extension time or use of alternative polymerases may be beneficial.
Q2: Can I use this kit for other tissue types besides tail?
A2: Yes. The kit is compatible with ear punches, toe clips, and other small tissue biopsies. Lysis time may require adjustment for denser or larger tissue samples.
Q3: How long can I store the crude lysate before PCR?
A3: Crude lysates may be stored at 4ˇăC for up to 1 week or at -20ˇăC for up to 1 month. Prolonged storage may result in DNA degradation and reduced PCR efficiency.
Q4: My PCR product shows smearing or non-specific bands. How can I improve specificity?
A4: Reduce the volume of crude lysate added to the PCR reaction, increase annealing temperature by 2-3ˇăC, or reduce the number of PCR cycles to 28-30 cycles.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of lysis and PCR conditions using control samples is strongly recommended before processing valuable experimental samples.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product and animal tissues.
Ordering Information
Catalog Number: IC-5561
Product Name: Mouse Tail Direct PCR Kit
Size: 50 reactions / 200 reactions / 500 reactions
Price:
50 reactions: CNY ¥800.00 / USD $80.00 / EUR €96.00 / JPY ¥14400.00
200 reactions: CNY ¥1900.00 / USD $190.00 / EUR €228.00 / JPY ¥34200.00
500 reactions: CNY ¥3800.00 / USD $380.00 / EUR €456.00 / JPY ¥68400.00
2023 Version
