Animal Tissue PCR Kit
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Price:
US$464.00
(Size: 100 rxns × 20 µl Systems)
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Abbexa's Animal Tissue PCR Kit uses a unique lysis buffer to lyse fresh or frozen animal tissues and blood. The resulting lysate without purification can be directly used as template for PCR amplification.
Kit Contents: Component | 100 rxns × 20 µl Systems | 500 rxns × 20 µl Systems |
AD1 Buffer | 4 ml | 20 ml |
AD2 Buffer | 1 ml | 5 ml |
AD3 Buffer | 4 ml | 2 × 10 ml |
2X PCR SuperMix (with dye) | 1 ml | 5 × 1 ml |
Nuclease-Free Water | 5 ml | 25 ml |
Target |
Animal Tissue PCR Kit |
Storage |
Store at -20 °C for up to two years. Avoid repeated freeze/thaw cycles. |
Buffer |
PCR SuperMix (with dye) contains ≤ 30% glycerol. The formulations of the AD1, AD2 and AD3 Buffers are proprietary. |
Availability |
Shipped within 10-20 working days. |
Note |
This product is for research use only. |
Directions for use |
Genomic DNA Extraction: - Immediately prior to carrying out the assay, prepare a AD1/AD2 Buffer mixture solution at a ratio of 4:1 (e.g. 40 µl of AD1 Buffer and 10 µl of AD2 Buffer) and mix thoroughly. 50 µl of AD1/AD2 Buffer mixture solution is sufficient for 1 sample/reaction. This mixture can be stored for up to 2 hours at room temperature.
- Prepare samples as follows:
- Mammalian cells - Recommended assay amount: ≤ 106 cells
Pellet cells by centrifugation and remove the supernatant. Add 50 µl of AD1/AD2 Buffer mixture solution to the pellet and mix thoroughly by pipetting up and down. - Saliva or Blood - Recommended assay amount: ≤ 10 µl
Add saliva or blood to 50 µl of AD1/AD2 Buffer mixture solution and mix thoroughly by pipetting up and down. - Animal tissues - Recommended assay amount: ≤ 10 mg (Mouse tail: ≤ 0.5 cm; Mouse ear: ≤ 0.5 cm2)
Cut tissue into small pieces using a sterile blade or scissors. Add 50 µl of AD1/AD2 Buffer mixture solution and mix thoroughly by pipetting up and down.
- Allow to stand at room temperature for 10 minutes (for samples that are hard to lyse such as hair tissue, the sample mixture can instead be incubated at 55 °C for 10 minutes), then incubate at 95 °C for 3 minutes.
- Add 40 µl of AD3 Buffer, then mix thoroughly. The lysate can be used as a PCR template, stored at 2-8 °C for up to 3 months, or stored at -20 °C for up to 6 months.
Reaction Components: Component | Volume | Final Concentration | Unpurified Lysate | Variable (≤ 4 µl) | as required | Forward Primer (10 µM) | 0.4 µl | 0.2 µM | Reverse Primer (10 µM) | 0.4 µl | 0.2 µM | 2X PCR SuperMix (with dye) | 10 µl | 1X | Nuclease-Free Water | Variable | N/A | Total Volume | 20 µl | N/A | Thermal Cycling Conditions: Number of Cycles | Temperature | Time per Cycle | 1 cycle | 94 °C | 5-10 min | 30-35 cycles | 94 °C | 30 seconds | 50-60 °C | 30 seconds | 72 °C | 1-2 kb/min | 1 cycle | 72 °C | 5-10 min | Notes: - Ensure that the sample and reagents have completely thawed and are mixed well prior to use in the assay.
- If faint bands are observed, increase the quantity of PCR template used or increase the number of PCR cycles (maximum 40 cycles).
- If non-specific amplification bands are observed, adjust the annealing temperature or reduce the quantity of PCR template used.
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