No products in the cart.
Product Information
| Product Name | Cat.No. | Spec. |
| Mouse Genotyping Kit | G3311-100T | 100 T |
| G3311-500T | 500 T |
Description
This kit contains the mouse tail, toes, ears and fast pyrolysis buffer used for genotyping of PCR Mix, pyrolysis products of the need for the extraction and purification, genome can be directly as a template for PCR amplification, easy operation, little injury in mice, you just need to trace amounts of tissue samples to achieve efficient genotyping, knockout detection, transgenic identification, etc.
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 Handling Conditions
Wet ice transportation; Mouse Genotyping Lysis Buffer A, store at 4℃; The remaining components are stored at -20℃; It is valid for 12 months.
Component
| 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 |
| Product Manual | 1 | ||
Assay Protocol
1. Template preparation
a) Preparation of lysate:
| Component | Lysate (single sample) |
| Mouse Genotyping Lysis Buffer Aa | 40 µL |
| Mouse Genotyping Lysis Buffer B | 10 µL |
a. Animal Tissue Lysis Buffer A can release white precipitation at low temperature. Incubate the Lysis Buffer at 37℃ for 2-3min before use until the white precipitation disappears, without affecting use.
b) Mix gently and centrifuge instantaneously；
c) A 1-3 mm tail tip, 2-3 mm diameter (perforated) mouse ear or 2-3 mm toe (without nail length) were taken, and the single sample lysate was added to the sample to be cleaved, incubated at 50℃ for 10 min and 95℃ for 3 min for full cleavage；
d) After centrifugation at 12000 rpm for 1 min, the lysed tissue block was centrifuged to the bottom of the tube, and the supernatant could be directly used as a template for PCR reaction. The supernatant can be transferred to another sterile EP and stored at -20℃ for a long time.
2. PCR reaction system is recommended (20 μL)：
| 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 amount of template should not exceed 1/10 of the reaction system.
b：The final primer concentration range is 0.5-1.0 μM, and the recommended primer concentration is 0.5 μM. Too little primer may lead to low or no amplification, and too much primer may lead to non-specific amplification.
3. PCR amplification conditions are recommended：
| 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. After each sampling, the sampler shall be flushed into 2% sodium hypochlorite solution repeatedly to avoid cross-contamination between different samples.
3. The tissue should be cut as far as possible so that the cleavage is sufficient.
4. Mouse Genotyping Lysis Buffer A and Mouse Genotyping Lysis Buffer B should be used in combination. Do not leave the Lysis Buffer for too long.
5. If the tissue block is difficult to crack, the cracking 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.
For Research Use Only!
Ver. No.: V1.0-202111
Product recommendation
|
Cat.No.
|
Product Name
|
Spec.
|
Operation
|
|---|
|
G3026-01
|
dNTP Mixture (10 mM each)
|
1 mL
|
|
|
G3026-05
|
dNTP Mixture (10 mM each)
|
5×1 mL
|
|
|
G3027-01
|
dNTP Mixture (2.5 mM each)
|
1 mL
|
|
|
G3027-05
|
dNTP Mixture (2.5 mM each)
|
5×1 mL
|
|
|
G3302-01
|
2 x Taq PCR Master Mix
|
1 mL
|
|
|
G3302-05
|
2 x Taq PCR Master Mix
|
5×1 mL
|
|
|
G3304-01
|
2 x Fast sTaq PCR Master Mix
|
1 mL
|
|
|
G3304-05
|
2 x Fast sTaq PCR Master Mix
|
5×1 mL
|
|
|
G3305-01
|
2 x Fast Pfus PCR Master Mix
|
1 mL
|
|
|
G3305-05
|
2 x Fast Pfus PCR Master Mix
|
5×1 mL
|
|
|
G3306-01
|
2 x Fast High Fidelity PCR Master Mix
|
1 mL
|
|
|
G3306-05
|
2 x Fast High Fidelity PCR Master Mix
|
5×1 mL
|
|
|
G3307-01
|
2 x LA PCR Master Mix
|
1 mL
|
|
|
G3307-05
|
2 x LA PCR Master Mix
|
5×1 mL
|
|
|
G3310-100T
|
Animal Tissue Direct PCR Kit
|
100 T
|
|
|
G3310-500T
|
Animal Tissue Direct PCR Kit
|
500 T
|
|
|
G3311-100T
|
Mouse Genotyping Kit
|
100 T
|
|
|
G3311-500T
|
Mouse Genotyping Kit
|
500 T
|
