Product Informations

• Goods Code BDAG0052
• Goods Name Mouse Tissue Direct Expansion TaqMan qPCR Kit
• Category 601
• Instruction Manual /files/uploads/instructions/2023/BDAG0052_小鼠组织直扩TaqMan qPCR试剂盒_说明书.pdf

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Product Details

Reagent components：

Component name	Item number
	BDAG0052-1ml	BDAG0052-5ml	BDAG0052-10ml
2×Mouse Tissue Direct TaqMan qPCR Mix	1 mL	5 mL	10 mL
Buffer TLB03	20 mL	100 mL	2,000 mL
ROX ReferenceⅠ (50×) *	40 µL	200 µL	400 µL
ROX ReferenceⅡ (50×) *	40 µL	200 µL	400 µL

*Note: ROX Reference is an optional reagent and is not included in this product. The usage methods for different models of Real-Time PCR instruments are as follows:
1. ROX Reference Ⅰ(50×) is a high-concentration ROX and is applicable to the following models of Real-Time PCR instruments:
- Applied Biosystems 7300 Real-Time PCR System(Thermo Fisher Scientific)；
- Applied Biosystems 7900 Real-Time PCR System(Thermo Fisher Scientific)；
- Step One Plus Real-Time PCR System (Thermo Fisher Scientific)。
2. ROX Reference II (50×) is a low-concentration ROX and is applicable to the following models of Real-Time PCR instruments:
- AppliedBiosystems 7500 Real-Time PCR System (Thermo Fisher Scientific)；
- AppliedBiosystems 7500 FAST Real-Time PCR System (Thermo Fisher Scientific)。
3. When used with the following models of Real Time PCR amplifiers, ROX Reference is not required:
Roche, Bio-Rad, TaKaRa, Hongshi, Yarui, Borui, Tianlong and other Real-Time PCR instruments that do not require the addition of ROX calibration dyes.


Product Description
This kit contains rapid release reagents for mouse tissue DNA and TaqMan qPCR 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 amplified by qPCR, enabling rapid genotyping of mice.
The 2×Mouse Tissue Direct TaqMan qPCR Mix in the kit is a 2× concentration premixed reagent, containing hot-start DNA polymerase, Mg2+, dNTP and an optimized buffer. It is easy to operate and reduces the possibility of contamination, enabling high-throughput detection. The qPCR reaction can be carried out simply by adding primer probes and templates to the reaction system. The protective agent added to the system enables Mix to maintain high activity even after repeated freeze-thaw cycles.

Product features
1. The kit does not require nucleic acid extraction or purification of mouse tissues; it can directly conduct qPCR reactions, saving time and effort.
Buffer TLB03 can complete the lysis of mouse tissues and release genomic DNA within 10 minutes.

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 Buffer TLB03 to each sample respectively, vortex to mix well, and then heat the samples at 95℃ for 10 minutes.
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
1) Preparation of reaction systems

Component name	Item number (µL per sample)		Final concentration
	Mix A	Mix B
2×Mouse Tissue Direct TaqMan qPCR Mix Forward Primer (10 μM) Reverse Primer (10 μM) Probe (10 μM) Template* PCR Grade H2O	10 0.5     0.5 0.4            2 To 20	25 1.0          1.0 0.4 5 To 50	1× 0.25 μM 0.25 μM 0.2 μM / /
Total volume/rxn	20	50

*Note: It is recommended that the amount of DNA template added be 10-20% of the total reaction volume.
2) Reaction procedure

Step	Temp.	Time	Cycle
Initial Denaturation	95℃	5 min	1 Cycle
Denaturation Annealing / Extension / Data Collection	95℃ 60℃	15 sec 30 sec	35~45 Cycles

Application Examples
qPCR detection of gene-edited mouse tails using this kit can all be well completed for amplification.

 

Precautions
1. The thawing of qPCR Mix should be carried out on ice and repeated freezing and thawing should be avoided.
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. Please avoid nuclease contamination of the sample, otherwise it may cause difficulties in the amplification reaction.
4. Please read this instruction carefully and use this product strictly in accordance with the operation procedures and the recommended dosage.