1. plasmid preparation
   1. mini prep recombinant vector and parent vector
   2. linearize vector
      1. 子主题 1
   3. analysis of linearized vector
2. yeast preparation
   1. medium preparation
      1. YPD plates and liquid medium
      2. RDB and RD top agar (for his selection)
      3. RDHB and RDH top agar ( for cell viability)
   2. pichia growth
      1. inocula from plate
      2. pichia cultrure
      3. transformation
         1. regeneration of pichia takes 4-6 days
   3. laccase expression
      1. Topic
      2. screen on MM plate to induce the expression of laccase
      3. Screen on BMMY plate + ABTS + CuSO4