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