How to Purify Taq

Taq polymerase is a heat-resistant enzyme that is used for research and medical diagnosis. Scientists extract taq from the taq polymerase DNA gene and utilize it during DNA sequencing polymerase chain reaction applications. During such instances, the DNA pattern is either studied or multiplied to create a DNA strand made up of nucleotides. When purifying taq, it is important to expose the extract to suggested temperatures at the correct time.

Things You'll Need

  • Measuring device
  • Celsius thermometer
  • Lauria Bertaini broth
  • Plastic containers (2)
  • IPIG
  • Tris-HC1
  • pH 7.9
  • Dextrose
  • EDTA
  • Detergent (no phosphorus)
  • Lysozyme
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Instructions

    • 1

      Place 5ml of Lauria Bertaini broth in a plastic container.

    • 2

      Place the taq polymerase extract in the broth-filled container and heat for 12 hours at 37 degrees Celsius.

    • 3

      Add 0.2-0.5mM of IPIG and continue heating at 37 degrees Celsius for another 12 hours.

    • 4

      Mix 50mM Tris-HC1, pH 7.9, 50 mM dextrose and 1mM EDTA together in a plastic container to create a buffer.

    • 5

      Wash the taq polymerase with detergent, place in the buffer and insert 4 mg/mL of lysozyme.

    • 6

      Place the container in a room temperature atmosphere for five minutes.

    • 7

      Place the substance in a 75 degrees Celsius atmosphere for 30 minutes.

    • 8

      Remove the purified taq polymerase extract and store as desired.

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