Team:HokkaidoU Japan/Notebook/September27
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<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/September24|September 24]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/September28|September 28]]</div></div> | <div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/September24|September 24]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/September28|September 28]]</div></div> | ||
+ | |||
+ | =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 |
Latest revision as of 12:44, 26 October 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