Uppsala-Sweden/19 August 2009

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(Difference between revisions)
(New page: {{Uppsala-Sweden Template}} ==Preparing competent cells== ''Continued since yesterday'' We harvested the seed cultures from yesterday and will later inoculate 300 ml of SOB media with on...)
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==Gel run==
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==Gel on Taq grad PCR with pirA BB primer==
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A gel run was performed to evaluate one of the PCR runs from yesterday
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I made a new gel of the Taq PCR yesterday.
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Setup: [[Media:Example.jpg]]
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[[Image:20090819-pirA_grad_taq-labelled.jpg]]
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Result: [[Image:Example.jpg]]
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--[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC)
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Another gel run was started to evaluate the other PCR performed yesterday.......
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==Gel on the Phusion PCR on adh2(Y) and pirA (BB & grad)==
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[[Image:20090819_pirA_oldp_phus_grad-labelled.jpg]]
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--[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC)
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==Gel on Taq grad PCR with pirA primer variations and Colony PCR==
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[[Image:20090819-colonypcr_pirA_tests-labelled.jpg]]
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--[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC)
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==Incubation of #3 adh2(Z)==
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#3 adh2(Z) in 5ml LB + amp
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incubated overnight @37°C and 250rpm
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--[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC)
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==Incubation of competent cells==
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1ml in 300ml of LB
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--[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC)
==Tansforming ADH2 Z.mobilis==
==Tansforming ADH2 Z.mobilis==

Revision as of 16:45, 19 August 2009




Contents

Preparing competent cells

Continued since yesterday

We harvested the seed cultures from yesterday and will later inoculate 300 ml of SOB media with one seed culter for harvest tomorrow. We also put three seed stocks in the -80 freezer for future use.


Gel on Taq grad PCR with pirA BB primer

I made a new gel of the Taq PCR yesterday.

20090819-pirA grad taq-labelled.jpg

--Karl.brune 16:45, 19 August 2009 (UTC)

Gel on the Phusion PCR on adh2(Y) and pirA (BB & grad)

20090819 pirA oldp phus grad-labelled.jpg

--Karl.brune 16:45, 19 August 2009 (UTC)

Gel on Taq grad PCR with pirA primer variations and Colony PCR

20090819-colonypcr pirA tests-labelled.jpg

--Karl.brune 16:45, 19 August 2009 (UTC)

Incubation of #3 adh2(Z)

  1. 3 adh2(Z) in 5ml LB + amp


incubated overnight @37°C and 250rpm --Karl.brune 16:45, 19 August 2009 (UTC)

Incubation of competent cells

1ml in 300ml of LB --Karl.brune 16:45, 19 August 2009 (UTC)

Tansforming ADH2 Z.mobilis

Continued since yesterday

The results form the "on column" digested ADH" was good with many colonies on the 50µl plate. We picked 7 of them and started a Taq evaluation PCR

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