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Cat. No.: HY-K0534

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Mouse Tissue Direct PCR Kit (with Dye)

MCE Mouse Tissue Direct PCR Kit (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.

Storage :
Lysis Buffer: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot and store at low temperature to avoid repeated freeze–thaw cycles.

Lysis Stop Solution, Direct PCR Master Mix (2×, with Dye): -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 15 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 particularly suitable for mouse genotyping and transgenic identification, and recommended for amplification of targets within 1 kb.

Features of MCE Mouse Tissue Direct PCR Kit (with Dye):

1. Fast and Convenient: One-step DNA preparation from mouse tissues; template preparation completed within 10–15 min, and PCR amplification within 1 h.

2. Low Sample Requirement: Only ~5 mg tissue or 1–5 mm mouse tail is sufficient.

3. High Specificity and Efficiency: Optimized Taq polymerase system with strong template affinity and high amplification specificity, ideal for genotyping and transgenic analysis.

4. User-Friendly Operation: The master mix contains tracking dyes, allowing PCR products to be directly loaded for electrophoresis.

5. High-Throughput Compatibility: Lysis and PCR can be performed in 96-well plates, suitable for large-scale mouse sample screening.

Protocol

General Protocol

1. Preparation of mouse tissue DNA template

Step	Procedure
Sample collection	Collect approximately 5-10 mg mouse tissue or 1-5 mm mouse tail and place it into a 1.5 mL microcentrifuge tube.
Tissue lysis	Add 90 μL Lysis Buffer, gently vortex to immerse the tissue, briefly centrifuge, and incubate at 95°C for 15 min.
Termination	Add 10 μL Lysis Termination Solution and mix gently.
Centrifugation	Centrifuge at 12,000 rpm for 2 min to pellet residual tissue debris.
DNA template	Transfer the supernatant to a new tube and use directly for PCR. Store at 4°C for 2-3 d or at -20°C for long-term storage.

Note: a. Finely mincing the tissue is recommended to improve lysis efficiency;
b. A 15 min incubation is sufficient for most PCR applications. For difficult-to-lyse samples or higher DNA yield, extend to 30 min;
c. Complete tissue lysis is not required; residual debris is removed by centrifugation.

2. PCR reaction mixture

Component	20 μL reaction
Direct PCR Master Mix (2×, with Dye)	10 μL
Forward Primer (10 μM)	0.8 μL
Reverse Primer (10 μM)	0.8 μL
Tissue lysate supernatant	1 μL
ddH₂O	To 20 μL

3. PCR cycling program

Step	Temperature	Time	Cycles
Initial denaturation	95°C	5 min	1
Denaturation	95°C	30 sec	30-35
Annealing	50-65°C	30 sec	30-35
Extension	72°C	30-60 sec/kb	30-35
Final extension	72°C	5 min	1

The Master Mix contains loading dye, so PCR products may be loaded directly onto an agarose gel after amplification.

Storage

Lysis Buffer: 4°C, 1 year. If not used for an extended period, it is recommended to aliquot and store at low temperature to avoid repeated freeze–thaw cycles.

Lysis Stop Solution, 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-K0534-50 T	Storage
HY-K0534-A	Mouse Tissue Direct PCR Reagent A	Lysis Buffer	1 mL × 5	4°C, 1 year.
HY-K0534-B	Mouse Tissue Direct PCR Reagent B	Lysis Termination Solution Direct PCR Master Mix (2×, with Dye)	0.6 mL 0.5 mL	-20°C, 1 year. Avoid repeated freeze-thaw cycles.