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Virus Isolation using the Minilys Personal Homogenizer

Instrument: Minilys Personal Homogenizer

Sample: Transfected HEK 293T producer cells

Protocol:

Minilys Method

  • 7mL tubes containing 1.4mm ceramic beads and using 5mL AAV resuspension buffer
  • 5000rpm, 2x60sec, 30sec breaks

Verses 4 freeze-thaw cycles method

Lysates from each method are used to transduce cultured HEK 293T and rAAV infectivity is compared by observation under the microscope.

Results:

The images show that the Minilys efficiently releases rAAV vectors on the resulting crude cell lysate is effectively used to transduce HEK 293T in image A. Transduction efficiency is similar compared to the freeze-thaw method in image B.