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Cat. No.: HY-K0535

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Mouse Tissue Direct PCR Plus Kit

MCE Mouse Tissue Direct PCR Plus Kit 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.

Storage :
Lysis Buffer: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot it appropriately for storage.

Proteinase K (20 mg/mL), Direct PCR Master Mix (2×): -20°C, 1 year. Avoid repeated freeze-thaw cycles.

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Use of MCE Products

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Description & Advantages

This kit utilizes an optimized lysis system, requiring only 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. It is suitable for mouse genotyping and transgenic identification.

Features of MCE Mouse Tissue Direct PCR Plus Kit:

1. Fast and Convenient: One-step DNA preparation from mouse tissues with high lysis efficiency; template DNA can be obtained within 15 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. High Specificity and Efficiency: Optimized Taq polymerase system with strong template affinity and high amplification specificity, ideal for genotyping and transgenic analysis.

4. High Amplification Efficiency: Capable of amplifying target fragments of varying sizes (1–6 kb), with strong GC compatibility, fast amplification speed, and high efficiency.

5. Multiplex PCR Capability: Supports up to 5-plex PCR amplification (200 bp–1000 bp).

6. Broad Sample Compatibility: Can also be extended to other sample types, such as HEK293 cells, saliva, and scalp tissue.

Protocol

General Protocol

1. Mouse tissue lysis

Step	Procedure
Sample collection	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 mixture	For each sample, mix 200 μL Lysis Buffer with 5 μL Proteinase K (20 mg/mL), then add to the tissue.
Tissue lysis	Mix thoroughly and incubate at 55°C for 10 min to release genomic DNA.
Inactivation	Incubate at 95°C for 5 min to inactivate Proteinase K and briefly centrifuge.
Template use	Use the supernatant as PCR template and avoid aspirating obvious tissue debris.

Note: Fresh tissues are recommended. Avoid repeated freeze-thawing of long-term frozen tissues. Sampling tools should be thoroughly cleaned between samples to prevent cross-contamination.

2. PCR reaction mixture

Component	50 μL reaction
Direct PCR Master Mix (2×)	25 μL
Forward Primer (10 μM)	2 μL
Reverse Primer (10 μM)	2 μL
Tissue lysate	1-5 μL
ddH₂O	To 50 μL

3. PCR cycling program

Step	Temperature	Time	Cycles
Initial denaturation	95°C	5 min	1
Denaturation	95°C	15 sec	30-35
Annealing	55-65°C	15-30 sec	30-35
Extension	72°C	10-30 sec/kb	30-35
Final extension	72°C	5 min	1

PCR products may be used for downstream agarose gel electrophoresis and genotyping analysis.

Storage

Lysis Buffer: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot it appropriately for storage.

Proteinase K (20 mg/mL), Direct PCR Master Mix (2×): -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-K0535-50 T	Storage
HY-K0535-A	Mouse Tissue Direct PCR Reagent Plus A	Lysis Buffer	12 mL	4°C, 1 year.
HY-K0535-B	Mouse Tissue Direct PCR Reagent Plus B	Proteinase K (20 mg/mL) Direct PCR Master Mix (2×)	625 μL 1.25 mL	-20°C, 1 year. Avoid repeated freeze-thaw cycles.