Product Informations

• Goods Code BDAG0026
• Goods Name Mouse Tissue One-Step Direct Amplification PCR Kit (With Dye)
• Category 601
• Instruction Manual /files/uploads/instructions/2024/202401/BDAG0026_小鼠组织一步法直扩 PCR 试剂盒(With Dye)_说明书.pdf

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Product Details

Item number	Specification	Number of reactions
BDAG0026-1ml	1ml	100rxn（10ul/rxn）
BDAG0026-5ml	5ml	500rxn（10ul/rxn）

Product Description
This kit contains rapid release reagents for mouse tissue DNA and PCR amplification reagents. It can rapidly release genomic DNA from tissues such as the tail, ears, and toes of mice without the need for nucleic acid extraction. The released products can be directly subjected to PCR amplification, enabling rapid genotyping of mice.
The Mouse Tissue Direct PCR Mix in the kit is a 2× concentration premix reagent, containing Taq DNA Polymerase, Mg2+, dNTP and an optimized buffer. It is easy to operate and reduces the possibility of contamination, enabling high-throughput detection. Just add primers and templates to the reaction system to carry out the PCR reaction. The protective agent added to the system enables the 2×Master Mix to remain highly active after repeated freeze-thaw cycles. The product contains dyes, and the PCR products can be directly subjected to electrophoresis detection.


Product features
1. The kit does not require nucleic acid extraction or purification of mouse tissues; it can directly conduct PCR reactions, saving time and effort
2. Mouse Tissue Lysis Buffer combined with proteinase K can rapidly lyse mouse tissues;
The best effect is achieved when the amplification product is within 2kb.
4. The product contains dyes, and the PCR products can be directly subjected to electrophoresis detection.

Experimental procedure
1. Sample preparation
Cut 1-5mm of the mouse tail (or 3-5mm diameter mouse ears and 2-3 mouse toes).
2. Add 200µL of Mouse Tissue Lysis Buffer and 4µL of Proteinase K to the samples respectively, vortex to mix well, and incubate ina 55 ° C water bath for 30 minutes. After incubation, heat the samples at 95 ° C for 10 minutes.
Note: If a large number of samples are to be processed, Mouse Tissue Lysis Buffer and Proteinase K can be mixed in advance, swirled evenly and set aside for later use.
3. Thoroughly mix the above products by vortex oscillation, then centrifuge at 12,000 RPM for 2 to 5 minutes. Take the supernatant and the PCR reaction can be carried out.

2. Amplification system and conditions
Preparation of the reaction system

Component	Volume(µL per sample)		Final concentration
	Mix A	Mix B
2×Mouse Tissue Direct PCR Mix Forward Primer (10 μM) Reverse Primer (10 μM) Template*  PCR Grade H2O	10 0.5     0.5    1 To 20	25 1.0          1.0 2.5 To 50	1× 0.25 μM 0.25 μM / /
Total volume/rxn	20	50

*Note: It is recommended that the amount of DNA template added be 5-10% of the total reaction volume

2. Reaction procedure

Step	Temp.	Time	Cycle
Initial Denaturation	95℃	2 min	1 Cycle
Denaturation Annealing*1 Extension*2 Final Extension	95℃ 60℃ 72℃ 72℃	25 sec 25 sec 1 min 7 min	35~40 Cycles

*Note: *1: The Annealing temperature is set according to the actual annealing temperature (Ta) of the primer.
The extension speed is 30sec/kb, which can be adjusted according to the size of the amplified fragment.

3. Detection of amplification products
   After PCR amplification is completed, the product can be directly detected by agarose gel electrophoresis.
   
   Application Examples
   This kit was used to detect genes of different fragment sizes in mouse tails, and amplification was all accomplished well.

Precautions
1. The thawing of PCR Mix needs to be carried out on ice and avoid repeated freezing and thawing.
2. The PCR reaction preparation process should be carried out on ice. After adding all the components, please briefly vortex to mix well and then centrifuge.
3. The inactivation step of Proteinase K is indispensable in the sample preparation stage.
4. Please avoid nuclease contamination of the sample, otherwise it may cause difficulties in the amplification reaction.
5. Please read this instruction carefully and use this product strictly in accordance with the operation procedures and the recommended dosage.