User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/10: 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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'''Golden gate trial 1''' | '''Golden gate trial 1''' | ||
* Add total 10.0 μL run to 50 μL DH5α-Turbo; incubate on ice 10 min.; plate on amp agar | * Add total 10.0 μL run to 50 μL DH5α-Turbo; incubate on ice 10 min.; plate on amp agar | ||
* Colonies: 7 on assembly plate, 9 on vector-only control. Pick 4 and make 2 mL liquid cultures. | |||
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PCR | PCR | ||
* 95°C, 3 min. | * 95°C, 3 min. | ||
* [95°C, | * [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35 | ||
* 72°C, 3 min. | * 72°C, 3 min. | ||
* 4°C, ∞ | * 4°C, ∞ | ||
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Dilutions | Dilutions: to final concentration of 20 fmol/μL | ||
{| {{table}} border="1" cellspacing="3" <!-- Dilution table --> | {| {{table}} border="1" cellspacing="3" <!-- Dilution table --> | ||
|- valign="top" | |- valign="top" | ||
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Golden Gate Reactions | |||
# hPCD + BL01 + pSB1A3 | # hPCD + BL01 + pSB1A3 | ||
# pSB1A3 | # pSB1A3 | ||
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|} | |} | ||
Thermal | Thermal cycling | ||
* [45°C, 2 min.; 16°C, 5 min.] x25 | * [45°C, 2 min.; 16°C, 5 min.] x25 | ||
* 60°C, 20 min. | * 60°C, 20 min. | ||
* 80°C, 20 min. | * 80°C, 20 min. | ||
* 4°C, ∞ | |||
Latest revision as of 22:20, 26 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
01/10/13
Violacein
Golden gate trial 1
Golden gate trial 2
PCR
Thermal cycling
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