Team:HokkaidoU Japan/Notebook/September27

From 2010.igem.org

Finally Salmonella's BAC clones have arrived!

  • BAC clones of S. Typhimurium, B_STM07H21 SGSC4024 1464540-1562427
    • Codes Type 3 Secretion Aparatus
  • BAC clones of S. Typhimurium, B_STM02P01 SGSC4014 837453-941863
    • Codes Secretion Signal



Culture of the BAC clone

  • made 25 ug/mL Chloramphenicol LB Agar plate
  • made 25 ug/mL Chloramphenicol LB medium

Preculture

  • while preparing the plate, cultibated each of the clones in LB medium for 2 hrs

Culture

  • plated the precultured BAC clones into LBC plate respectively



Electroporation

Purification

In order to decrease salt concentration, purified 1 ng/uL of BAC solution which has been prepared on Thursday using Microcon YM-10


Electroporation

electroporated DH5a and MG1655 using 1 uL of purified DNA solution

  • MG1655: 2.5 kV: 5.4 ms
  • DH5a: 2.49 kV: 5.7 ms
  • added 200 uL of SOB and transfered into round tube
  • incubated at 37C for 2 hrs
  • plated onto LBA