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

37.1. Date#

15072020

37.2. Objective#

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

37.3. 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

37.3.1. Sanity check#

  • 07082020- Plate 12 colonies of ylic133+pBk549 for the sanity check

  • Colony #10 and #12 passed the test

37.4. Results#

  • I have plenty of colonies in the transformation plate :)

  • None from the negative control. :)

  • Proceed with these two colonies for SATAY