Uppsala-Sweden/6 September 2009

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==Moving ON to fride==
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{{Uppsala-Sweden Template}}
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ON cultures for the lux system were moved to the iGEM fridge for MiniPrep tomorrow
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==Gels of Colony PCR==
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==Checking Plates==
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[[Image:20090906-colonypcr_gel1_40-labelled.jpg]]
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All plates show colonoies.
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[[Image:20090906-colonypcr_gel2_40-labelled.jpg]]
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After picking colonies -> fridge.
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--[[User:Karl.brune|Karl.brune]] 23:08, 5 September 2009 (UTC)
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==Picking Colonies==
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[[Image:20090906-colonypcr_gel3_40-labelled.jpg]]
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I picked colonies from plates into 10µl selective media.
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==Inoculation==
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--[[User:Karl.brune|Karl.brune]] 23:08, 5 September 2009 (UTC)
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5 Colonies of pirA (AC3) -> pirA-X
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5 Colonies of b0034-pirA (AC3) -> RA-X
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5 Colonies of B0034-Yadh2-B0015 (AC3) -> RYB-X
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5 Colonies of B0015 (AC3) - T-X
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5 Colonies of R0011-B0034 (AK3) -> RR-X
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3 Colonies of E1A1-B0015 (AK3) -> E1A1X
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Promising culutures according to the gels were inoculated in their specific antibiotic.
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3 Colonies of E1A2-B0015 (AK3) -> E1A2X
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3 Colonies of E1A3-B0015 (AK3) -> E1A3X
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3 Colonies of E1A4-B0015 (AK3) -> E1A4X
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3 Colonies of E1B1-B0015 (AK3) -> E1B1X
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'''Subunit E2'''
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3 Colonies of E1B2-B0015 (AK3) -> E1B2X
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#E21-1
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3 Colonies of E1B3-B0015 (AK3) -> E1B3X
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#E22-3
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3 Colonies of E1B4-B0015 (AK3) -> E1B4X
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#E23-3
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#E24-1
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3 Colonies of E21-B0015 (AK3) -> E21X
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'''Subunit E1 alpha'''
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3 Colonies of E22-B0015 (AK3) -> E22X
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#E1A1-1
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3 Colonies of E23-B0015 (AK3) -> E23X
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#E1A1-2
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3 Colonies of E24-B0015 (AK3) -> E24X
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#E1A2-2
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#E1A2-3
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#E1A3-1
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#E1A4-2
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X= # of colony
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'''Subunit E1 beta'''
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#E1B1-3
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#E1B2-3
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#E1B3-1
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#E1B4-2
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--[[User:Karl.brune|Karl.brune]] 23:08, 5 September 2009 (UTC)
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'''B0015, B0034-pirA, R0011-B0034, B0034-Yadh2-B0015'''
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#T-1
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#RP-2
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#RR-4
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#RR-5
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#RYT-1
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#RYT-2
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==Colony PCR==
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Furthermore these were inoculated (Lux System etc.)
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TAQ colony PCR was performed.
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'''A340620, C0621, J237034'''
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#A340620
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#C0621
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#J237034
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10µl reaction, standart mix with verfication primer VF2 and VR.
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'''and these here:'''
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#R0062 (A)
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Initial Denat: 1min 30sec @94°C
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#RARP 4, 5 ,7 (CH)
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40Cylces
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Denaturation: 45 sec @94°C
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Annealing: 45 sec @50°C
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Elongation: 1min 45sec @ 72°C
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Return to Denat.
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Final Elongation: 10min @72°C
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Infinite storage at 4°C
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Colony tubes were moved to fridge after taking template for the PCR
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--[[User:Karl.brune|Karl.brune]] 23:08, 5 September 2009 (UTC)
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Latest revision as of 19:03, 6 September 2009




Gels of Colony PCR

20090906-colonypcr gel1 40-labelled.jpg

20090906-colonypcr gel2 40-labelled.jpg

20090906-colonypcr gel3 40-labelled.jpg

Inoculation

Promising culutures according to the gels were inoculated in their specific antibiotic.

Subunit E2

  1. E21-1
  2. E22-3
  3. E23-3
  4. E24-1

Subunit E1 alpha

  1. E1A1-1
  2. E1A1-2
  3. E1A2-2
  4. E1A2-3
  5. E1A3-1
  6. E1A4-2

Subunit E1 beta

  1. E1B1-3
  2. E1B2-3
  3. E1B3-1
  4. E1B4-2

B0015, B0034-pirA, R0011-B0034, B0034-Yadh2-B0015

  1. T-1
  2. RP-2
  3. RR-4
  4. RR-5
  5. RYT-1
  6. RYT-2

Furthermore these were inoculated (Lux System etc.)

A340620, C0621, J237034

  1. A340620
  2. C0621
  3. J237034

and these here:

  1. R0062 (A)
  2. RARP 4, 5 ,7 (CH)
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