User:Karmella Haynes/Notebook/BioBrick cloning/2012/11/30: 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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| colspan="2"| | | colspan="2"| | ||
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---- | ---- | ||
'''Assemblies''' | '''Assemblies''' | ||
# KAH3/MV2: (1) KAH3/(X/P)/751 + (2) MV2/( | # KAH3/MV2: (1) KAH3/(X/P)/751 + (2) MV2/(S/P)/4529 | ||
> Digests (Fermentas FD)<br> | > Digests (Fermentas FD)<br> | ||
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|-valign="top" | |-valign="top" | ||
| <u>Reagent</u> || <u>Volume</u> || | | <u>Reagent</u> || <u>Volume</u> || | ||
| rowspan="9" | [[Image: | | rowspan="9" | [[Image:KAH113012_gel1.jpg |150px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
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| DNA (plasmid) || up to 25 μL | | DNA (plasmid) || up to 25 μL | ||
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| <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u> | | <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u> | ||
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| 1. KAH3 (X/P) || | | 1. KAH3 (X/P) || 0.028 || 1.96 || 28.41 | ||
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| 2. MV2 (S/P) || | | 2. MV2 (S/P) || 0.085 || 1.93 || 85.02 | ||
|} | |} | ||
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| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 11/31/12</font> | | <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 11/31/12</font> | ||
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| 1. KAH3 (X/P)/751, 8 ng + MV2 (S/P)/4529, 25 ng || <font color="blue">KAH3/MV2 | | 1. KAH3 (X/P)/751, 8 ng + MV2 (S/P)/4529, 25 ng || <font color="blue">KAH3/MV2 >10:1 (Pick 2)</font> | ||
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| 2. MV2 (S/P)/ 25 ng || | | 2. MV2 (S/P)/ 25 ng || zero colonies | ||
|} | |} | ||
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| || 1 || 2 || | | || 1 || 2 || | ||
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| Insert DNA || | | Insert DNA || 0.3 || --- || | ||
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| Vector DNA || | | Vector DNA || 0.3 || 0.3 || | ||
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| 2x lgn buf (Roche) || 5.0 || 5.0 || | | 2x lgn buf (Roche) || 5.0 || 5.0 || | ||
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| T4 ligase (NEB) || 1.0 || 1.0 || | | T4 ligase (NEB) || 1.0 || 1.0 || | ||
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| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 3.4 || 3.7 || | ||
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| || 10 μL || 10 μL || | | || 10 μL || 10 μL || | ||
|} | |} | ||
* Follow quick transformation protocol | |||
* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar | |||
Latest revision as of 22:18, 26 September 2017
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11/30/12
Assemblies
> Digests (Fermentas FD)
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