Naegleria 5

8 QIAprep Spin miniprep kits for 4 cultures each

Notes before starting

• Optional: Add LyseBlue reagent to Buffer P1 at a ratio of 1 to 1000.
• Add the provided RNase A solution to Buffer P1, mix and store at 2–8°C.
• Add ethanol (96–100%) to Buffer PE before use (see bottle label for volume).
• All centrifugation steps are carried out at 13,000 rpm (~17,900 x g) in a conventional table-top microcentrifuge.

Each kit contains:

• 4 spin columns
• 6 buffers, as below

Other materials

• Age1 + cutsmart
• 8 2mL tubes of LB broth
• competent HT115 cells from -80
• two 1% gels for wide gel rig, 2 small-well combs each
  • 2 g agarose
  • 200 mL 1X TAE
  • 20 uL SYBR Safe
• 1X TAE running buffer
• 1KB DNA standard

Equipment

• Microcentrifuges capable of 13000rpm
• Hot blocks at
  • 37
  • 42
• two wide gel rigs
• two power supplies
• two blue light boxes with photography box