User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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| DNA(plasmid) || 0.2 μL | | DNA(plasmid) || 0.2 μL | ||
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| primer 1 || 1.0 | | 10 μM primer 1 || 1.0 | ||
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| primer 2 || 1.0 | | 10 μM primer 2 || 1.0 | ||
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| 2x GoTaq || 25.0 | | 2x GoTaq || 25.0 | ||
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Transformation | Transformation | ||
* | * Transfer 10 μL reactions into 2.0 mL round bottom tubes; add 50 μL chemically competent BL21 cells (thawed on ice); pipette up and down 3x to mix | ||
* Incubate on ice for 5 min. | |||
* Heat shock at 42°C (heat block) for exactly 45 sec.; place on ice immediately | * Heat shock at 42°C (heat block) for exactly 45 sec.; place on ice immediately | ||
* Add 750 μL plain LB broth; 37°C incubator: lay tubes flat and tape to the shaking rack; incubate with shaking for 45 min. | * Add 750 μL plain LB broth; 37°C incubator: lay tubes flat and tape to the shaking rack; incubate with shaking for 45 min. |
Latest revision as of 22:29, 26 September 2017
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02/23/13
Type IIS Assembly, PCR: LOV-H2B+his
PCR
Golden Gate Reactions
Thermal cycling
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