Animal Tissue PCR Kit

Catalogue No: abx098011
Price: £230.00
(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 Buffer4 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.
Directions for use Genomic DNA Extraction:
  1. 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.
  2. 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.
  3. 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.
  4. 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 °C5-10 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.
Availability Shipped within 10-20 working days.
Note This product is for research use only.
Research Articles on Animal Tissue PCR Kit

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