Naegleria 5

Naegleria 5 kdorfman Tue, 02/08/2022 - 21:08

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
Buffer uL per rxn mL per 4.5 rxns
P1 250 1.125
P2 250 1.125
N3 350 1.575
PB 500 2.25
PE 750 3.375
EB 50 0.225

Other materials

Equipment

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