Uppsala-Sweden/19 August 2009
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(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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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. | 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. | ||
+ | ==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 | |
- | + | ==Gel on Taq grad PCR with pirA BB primer== | |
- | + | I made a new gel of the Taq PCR yesterday. | |
- | + | [[Image:20090819-pirA_grad_taq-labelled.jpg]] | |
- | + | --[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC) | |
+ | ==Gel on the Phusion PCR on adh2(Y) and pirA (BB & grad)== | ||
- | + | [[Image:20090819_pirA_oldp_phus_grad-labelled.jpg]] | |
- | + | ||
- | + | --[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC) | |
+ | |||
+ | ==Gel on Taq grad PCR with pirA primer variations and Colony PCR== | ||
+ | |||
+ | [[Image:20090819-colonypcr_pirA_tests-labelled.jpg]] | ||
+ | |||
+ | --[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC) | ||
+ | |||
+ | ==Incubation of #3 adh2(Z)== | ||
+ | |||
+ | #3 adh2(Z) in 5ml LB + amp | ||
+ | |||
+ | |||
+ | incubated overnight @37°C and 250rpm | ||
+ | --[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC) | ||
+ | |||
+ | ==Incubation of competent cells== | ||
+ | |||
+ | 1ml in 300ml of LB | ||
+ | --[[User:Karl.brune|Karl.brune]] 16:45, 19 August 2009 (UTC) |
Latest revision as of 16:48, 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.
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
Gel on Taq grad PCR with pirA BB primer
I made a new gel of the Taq PCR yesterday.
--Karl.brune 16:45, 19 August 2009 (UTC)
Gel on the Phusion PCR on adh2(Y) and pirA (BB & grad)
--Karl.brune 16:45, 19 August 2009 (UTC)
Gel on Taq grad PCR with pirA primer variations and Colony PCR
--Karl.brune 16:45, 19 August 2009 (UTC)
Incubation of #3 adh2(Z)
- 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)