Mouse Tissue Direct PCR Kit – Pro (with Dye)
Based on 1 Customer Validation
MCE Mouse Tissue Direct PCR Kit – Pro (with Dye) is specifically designed for direct PCR amplification from tissue samples. It enables rapid, one-step release of genomic DNA from various mouse tissues (such as tail, ear, toe, and muscle), which can be directly used for downstream PCR amplification and analysis without the need for conventional DNA extraction and purification steps, thereby significantly simplifying the experimental workflow.
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Storage :
Lysis Buffer, Lysis Termination Solution: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot it appropriately for storage.
Direct PCR Master Mix (2×, with Dye): -20°C, 1 year. Avoid repeated freeze-thaw cycles.
- Manual (5787KB)
- COA
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Mouse Tissue Direct PCR Kit – Pro (with Dye) is specifically designed for direct PCR amplification from tissue samples. It enables rapid, one-step release of genomic DNA from various mouse tissues (such as tail, ear, toe, and muscle), which can be directly used for downstream PCR amplification and analysis without the need for conventional DNA extraction and purification steps, thereby significantly simplifying the experimental workflow.
This kit utilizes an optimized lysis system, requiring only 5-10 min for tissue digestion to generate PCR-ready DNA templates. In addition, the provided PCR Master Mix contains all essential components for PCR amplification, including Taq DNA polymerase, dNTPs, reaction buffer, and loading dye. Users only need to add the template and primers to perform amplification. The PCR products can be directly analyzed by agarose gel electrophoresis without the need for additional loading buffer, effectively reducing operational complexity and the risk of contamination. It is suitable for mouse genotyping and transgenic identification.
Features of MCE Mouse Tissue Direct PCR Kit – Pro (with Dye):
1. Fast and Convenient: One-step DNA preparation from mouse tissues with high lysis efficiency; template DNA can be obtained within 10 min.
2. Low Sample Requirement: Only 2-5 mm2 mouse ear or liver tissue, 1–5 mm mouse tail, or 1–2 mouse toes are sufficient for analysis.
3. Flexible Lysis Conditions: Supports two lysis options—95°C for 3 min or 60°C for 15 min—both enabling efficient DNA release.
4. Stable Lysates: The lysates can be stored at 25°C or 4°C for up to 4 weeks and still be used for downstream PCR amplification.
5. High Specificity and Efficiency: Optimized Taq polymerase system with strong template affinity and high amplification specificity, ideal for genotyping and transgenic analysis.
6. High Amplification Efficiency: Efficient amplification of target fragments up to ≤ 10 kb, with strong GC compatibility and fast extension rates (5–10 kb/s), enabling time-efficient workflows.
7. Multiplex PCR Capability: Supports 5–7 plex PCR amplification (500 bp–5 kb).
8. Broad Sample Compatibility: In addition to tissue samples, compatible with various sample types, including blood and cell suspensions.
9. High Kit Stability: The complete kit can be stored at 25°C or 4°C for up to 4 weeks with stable performance, meeting experimental requirements.
Protocol
General Protocol
1. Preparation of mouse tissue DNA template
| Lysis option | Procedure |
|---|---|
| Sample | Collect 2-5 mm2 mouse ear/liver tissue, 1-5 mm mouse tail, or 1-2 mouse toes and place into a centrifuge tube. |
| Lysis | Add 50 μL Lysis Buffer and mix gently to ensure the tissue is immersed. |
| Rapid lysis | Incubate at 95°C for 3 min. |
| Mild lysis | Incubate at 60°C for 15 min. |
| Termination | Add 50 μL Lysis Termination Solution and mix gently. The lysate may be used directly for PCR. |
Note: Either lysis condition can efficiently release DNA and may be selected according to sample type and workflow. The resulting lysate may be stored short-term at 25°C or 4°C.
2. PCR reaction mixture
| Component | 50 μL reaction |
|---|---|
| Direct PCR Master Mix (2×, with Dye) | 25 μL |
| Forward Primer (10 μM) | 2 μL |
| Reverse Primer (10 μM) | 2 μL |
| Tissue lysate | 2-5 μL |
| ddH2O | To 50 μL |
3. PCR cycling program
| Step | Temperature | Time | Cycles |
|---|---|---|---|
| Initial denaturation | 95°C | 3 min | 1 |
| Denaturation | 95°C | 10 sec | 30-35 |
| Annealing | 55-65°C | 10-15 sec | 30-35 |
| Extension | 72°C | 5-10 sec/kb | 30-35 |
| Final extension | 72°C | 5 min | 1 |
The kit is suitable for rapid amplification and multiplex PCR. The Master Mix contains loading dye, so PCR products may be loaded directly onto an agarose gel.
Storage
Lysis Buffer, Lysis Termination Solution: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot it appropriately for storage.
Direct PCR Master Mix (2×, with Dye): -20°C, 1 year. Avoid repeated freeze-thaw cycles.
Attention
1. To prevent cross-contamination between samples, sampling tools should be thoroughly cleaned after each use. It is recommended to immerse the cutting edge or any part in direct contact with the sample in 2% sodium hypochlorite solution, rinse repeatedly, and then wipe dry with clean tissue before reuse. For improved efficiency, multiple sets of sampling tools may be prepared and cleaned collectively to ensure that each sample is handled with contamination-free instruments.
2. Freshly collected animal tissues are recommended for optimal results. For long-term frozen samples, repeated freeze–thaw cycles should be avoided, as they may lead to genomic DNA degradation and consequently affect PCR amplification efficiency and data reliability.
3. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Cat. No. | Product List | Components | HY-K0536-50 T | Storage |
|---|---|---|---|---|
| HY-K0536-A | Mouse Tissue Direct PCR Reagent Pro A | Lysis Buffer Lysis Termination Solution |
1.25 mL × 2 1.25 mL × 2 |
4°C, 1 year. |
| HY-K0536-B | Mouse Tissue Direct PCR Reagent Pro B | Direct PCR Master Mix (2×, with Dye) | 1.25 mL | -20°C, 1 year. Avoid repeated freeze-thaw cycles. |
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