Process Flow of PCR Analysis with DNA Test Kits – Summary

Extraction

Summary

  • Animal sample (Should be less than 10 mg.)
  • GenCheck 40 ul
  • High speed Centrifuge at 10,000 rpm for 30 seconds.
  • Thermo-shaker at 98-100 ° C for 10 minutes.
  • Stay on ice for 1 minute.
  • Centrifuge at 10,000 rpm for 5 minutes.
  • Transfer 5μl of the supernatant and add 95μl of RNase-free water.

Amplification

For Sample

  • PCR enzyme Primer 5μl per sample
  • RNase-free water 2μl per sample
  • Primer Solution 2μl per sample
  • Extracted DNA 1μl
  • Micropipette
  • Centrifuge
  • Thermocycler

For Positive Controls and Negative Controls:

  • PCR enzyme Primer 5 ul x (Number of Positive Controls available in the Test Kit)
  • RNase-free Water 2μl x (Number of Positive Controls available in the Test Kit)
  • Primer Solution 2μl x (Number of Positive Controls available in the Test Kit)
  • For Positive Control(s): Put 1μl of Positive Control for each disease (EHP, WSSV, IHHNV, AHPND/EMS).
  • For Negative Control: Put 1μl of RNAse-Free Water.
  • Micropipette
  • Microcentrifuge (using Multi Spin low speed)
  • Thermocycler (ASTEC 325 GeneAtlas PCR machine or other equipment recommended).

Identification

For Sample

  • Latex Solution 1.5μl
  • Dilution Buffer 20μl
  • Positive Controls 1μl
  • Amplified DNA 1μl
  • Micropipette
  • C-PAS Strip (1 Strip per sample)