Naegleria 3

Submitted by kdorfman on Sat, 12/18/2021 - 12:00

Cloning Bootcamp

  • 37C heatblock
  • 42C heatblock
  • Age1
  • 10X buffer NEB1.1
  • rSAP
  • Sterile water for cloning
  • Ligase + 2X rapid lligase buffer
  • 1 sample: PCR cleanup kit (Qiaquick) (one per group)
  • 1 tube (5 rxns) Competent cells (DH5 alpha, just for cloning)
  • 5x LB+Amp plates
  • Sterile beads for spreading
  • 30 ng/ul backbone plasmid for cutting (estimated 15n/ul after elution w/ 30ul)
  • Gel purified inserts (gamma tub, centrin, sas-6 at 15ng/ul)
  • LB (plain) for transformation recovery