Team:Queens Canada/freeze

Freeze-Thaw Cycles Bacterial Lysis · Benchling

Freeze-Thaw Cycles Bacterial Lysis

Introduction

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Materials

    • Buffer
    • Lysozyme and EDTA

Procedure

  1. Resuspend a gram of cells in 1-2 mL buffer and incubate with 1 mg/mL lysozyme with 0.5 M EDTA in ice for 1 hour.
  1. Freeze the cells using liquid nitrogen and thaw at ROOM TEMPERATURE.
  • Repeat 5-8 times.
  1. Add DNAse and incubate at RT for 10-15 minutes and proceed to clarify lysate by centrifugation.