Page History

Description: 

Making HEK293 expressing LilrB2 and PirB on the cell membrane. Will test membrane localization by using anti-LilrB2 and anti-PirB antibodies in Experiment 2.

Plan:

•  Order LilrB2 and PirB gBlocks 
•  Simulate Golden Gate on Geneious and make sure it works 
•  Golden Gate gBlocks into ggDONR
•  Transform bacteria with Golden Gate product and plate 
•  Pick colonies and grow in liquid culture
•  Miniprep cells 
•  LR into pDEST

Status:

• First and second set of liquid cultures didn't grow, possibly because:
  • Too much media
  • Plates were old, antibiotic possibly not working
• First set of plates didn't have any blue colonies
• Second set of plates grew blue and white colonies
  • PirB didn't grow any white colonies
  • Colonies picked for LilrB2
    • Liquid culture grew
    • Miniprepped - Product to be verified
• Golden Gate repeated
  • cells transformed and plated
  • +ve control didn't grow
    • LilrB2 grew blue and white colonies
    • PirB grew white colonies
  • Both liquid cultures grew
  • Miniprepped - Product to be verified