Products
Molecular Biology Reagents | Consumables
Nucleic Acid Amplification
PCR Series
Mouse Genotyping Kit (for PCR series)
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Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
Mouse Genotyping Kit | G3311-100T | 100 T |
G3311-500T | 500 T |
Product Description/Introduction
This kit contains rapid lysis buffer for the mouse tail, toes, ears and PCR Mix for genotyping.The lysis product does not require genome extraction and purification, and can be used directly as a template for PCR amplification. It is easy to operate, causes little damage to mice, and requires only a trace amount of tissue samples to achieve highly efficient genotyping, knockout detection, and transgene identification.
The 2×Mouse Genotyping PCR Mix (+Dye) includes high-inhibition and rapid amplification of DNA polymerases, dNTPs, and optimized buffer system for high efficiency and specificity of direct amplification. It is suitable for target gene amplification up to 2 Kb.
Storage and Shipping Conditions
Ship with wet ice;Mouse Genotyping Lysis Buffer A store at 4; The remaining components are stored at -20, valid for 12 months.
Product Contents
Component Number | Component | G3311-100 | G3311-500 |
G3311-1 | Mouse Genotyping Lysis Buffer A | 4 mL | 20 mL |
G3311-2 | Mouse Genotyping Lysis Buffer B | 1 mL | 5 mL |
G3311-3 | 2×Mouse Genotyping PCR Mix (+Dye) | 1 mL | 5×1 mL |
Manual | One copy | ||
Assay Protocol / Procedures
Template Preparation
1. Prepare Lysis Solution
Component | Lysate (single sample) |
Mouse Genotyping Lysis Buffer Aa | 40 µL |
Mouse Genotyping Lysis Buffer B | 10 µL |
a:Mouse Genotyping Lysis Buffer A appear white precipitates under low temperature. Incubate at 37 for 2-3 min until the white precipitate disappears before use, it will not affect the use.
2. Mix gently and centrifuge instantaneously.
3. Take a 1-3 mm tail tip,a2-3 mm diameter (perforated) mouse ear ora2-3 mm toe（ excluding the length of the nail),add single-sample lysate to the sample to be lysed, incubate at 50°C for 10 minutes,and 95°C for 3 min for full lysis.
4. Centrifuge the lysed tissue block at 12000 rpm for 1 min to the bottom of the tube, and the supernatant can be used directly as a template for PCR reaction. The supernatant can be transferred to another sterile EP and placed at -20°C for long-term storage.
Recommended PCR reaction system (20μL):
Component | 20 μL rxn | Final Concentration |
Templatea | 1-2 μL | As required |
Forward Primer (10 μM)b | 1 μL | 0.5 μM |
Reverse Primer (10 μM)b | 1 μL | 0.5 μM |
2×Mouse Genotyping PCR Mix (+Dye) | 10 μL | 1× |
ddH2O | Add to 20 μL |
a：The dosage of template shall not exceed 1/10 of the reaction system.
b：Primer final concentration rang 0.5-1.0 μM. The recommended primer concentration is 0.5 μM. Too few primers may lead to low amplification yield or no amplification, and too many primers may lead to non-specific amplification.
5. Recommended PCR amplification conditions:
Step | Temperature | Time | Cycles |
Initial Denaturation | 98 | 2 min | 1 |
Denaturation | 98 | 15 s | 35 |
Annealing | 50-65 | 20 s | |
Extension | 72 | 10 s/kb | |
Final extension | 72 | 5-10 min | 1 |
Hold | 4-16 | Forever |
Note
1. Before using the reagent, please gently mix it upside down. Do not swirl and oscillate to avoid bubbles.
2. The sampler shall be flushed into 2% sodium hypochlorite solution repeatedly after collect samples to avoid cross-contamination between different samples.
3. The tissue should be cut into tiny piece so that the cleavage is sufficient.
4. Mouse Genotyping Lysis Buffer A and Mouse Genotyping Lysis Buffer B should be used as they are prepared, and should not be left for too long after mixing, otherwise the lysis effect will be affected.
5. If the tissue block is difficult to lysis, the lysi time can be extended to 20 min at 50.
6. After lysis, incomplete lysis tissue remains in EP tube, which is a normal phenomenon and does not affect the use.
