# Title : Transformation of ylic133 (WT ade- ura-) with pbK549 for SATAY

# Date

15072020

# Objective

• To integrate the WT in the SATAY round with ylic135 and ylic136

# Method

• Yeast Transformation Protocol

• 15072020: Incubation from glycerol stock in 10ml YPD+6xade

• 16072020: OD measurement : 11.3 at 9:35 am , so I did 20X dilution of the original culture to have around OD=0.5

  • Transformation stopped

• 23072020 Re-do of the transformation

  • at 9:35am OD measurement, OD=~6 , I diluted 10x times to a final volume of 10mL with YPD+6xade
  • at 11:55am OD measurement, OD=1,8 😃 So , the cells were ready for transformation.

• 3ul plasmid (46,7ng/ul) implies 140ng plasmid.

• I did one negative controls.

• Plating 150ul cells+50ul MiliQ and 50ul cells+150ul MiliQ in -URA+6xADE and 200ul cells of the negative control in -URA+6xade

# Sanity check

• [x] 07082020- Plate 12 colonies of ylic133+pBk549 for the sanity check
• Colony #10 and #12 passed the test

# Results

• I have plenty of colonies in the transformation plate 😃
• None from the negative control. 😃
• Proceed with these two colonies for SATAY