Team:HokkaidoU Japan/Notebook/September27

From 2010.igem.org

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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>
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=Finally Salmonella's BAC clones have arrived!=
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* BAC clones of S. Typhimurium, B_STM07H21  '''SGSC4024'''  1464540-1562427
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** Codes Type 3 Secretion Aparatus
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* BAC clones of S. Typhimurium, B_STM02P01　'''SGSC4014'''　837453-941863
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** Codes Secretion Signal
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 +
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----
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==Culture of the BAC clone==
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* made 25 ug/mL Chloramphenicol LB Agar plate
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* made 25 ug/mL Chloramphenicol LB medium
 +
 +
===Preculture===
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* while preparing the plate, cultibated each of the clones in LB medium for 2 hrs
 +
 +
===Culture===
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* plated the precultured BAC clones into LBC plate respectively
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 +
 +
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----
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==Electroporation==
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===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
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* 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
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* plated onto LBA

Latest revision as of 12:44, 26 October 2010

Notebook

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
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/September27"