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Team:HokkaidoU Japan/Notebook/September27
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| - | =Finally Salmonella | + | =Finally Salmonella's BAC clones have arrived!= |
* BAC clones of S. Typhimurium, B_STM07H21 '''SGSC4024''' 1464540-1562427 | * BAC clones of S. Typhimurium, B_STM07H21 '''SGSC4024''' 1464540-1562427 | ||
** Codes Type 3 Secretion Aparatus | ** Codes Type 3 Secretion Aparatus | ||
Latest revision as of 12:44, 26 October 2010
since 13 Jul 2010
since 1 Aug 2010
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
