User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/20: 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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| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/21/13</font> | | Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/21/13</font> | ||
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| 1. A1 (ds oligo S/S)/76, ''0.5 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 1. A1 (ds oligo S/S)/76, ''0.5 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 2. A1 (ds oligo S/S)/76, ''1.0 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 2. A1 (ds oligo S/S)/76, ''1.0 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 3. A1 (ds oligo S/S)/76, ''2.0 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 3. A1 (ds oligo S/S)/76, ''2.0 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 4. MV9(X dp)/ 25 ng || | | 4. MV9(X dp)/ 25 ng || | ||
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| 5. A3 (ds oligo S/S)/76, ''0.5 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 5. A3 (ds oligo S/S)/76, ''0.5 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 6. A3 (ds oligo S/S)/76, ''1.0 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 6. A3 (ds oligo S/S)/76, ''1.0 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 7. A3 (ds oligo S/S)/76, ''2.0 μL'' + MV2(X dp)/4529, 25 ng || <font color=" | | 7. A3 (ds oligo S/S)/76, ''2.0 μL'' + MV2(X dp)/4529, 25 ng || <font color="red">MV9 failed</font> | ||
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| 8. MV9(X dp)/ 25 ng || | | 8. MV9(X dp)/ 25 ng || | ||
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| 9. MV9(X no dp)/ 25 ng + T4 ligase || | | 9. MV9(X no dp)/ 25 ng + T4 ligase || <font color="blue">Re-ligation successful, ~40 colonies</font> | ||
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| 10. MV9(X no dp)/ 25 ng || | | 10. MV9(X no dp)/ 25 ng || | ||
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| Insert DNA || 0.5 || 1.0 || 2.0 || --- || 0.5 || 1.0 || 2.0 || --- || --- || --- | | Insert DNA || 0.5 || 1.0 || 2.0 || --- || 0.5 || 1.0 || 2.0 || --- || --- || --- | ||
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| Vector DNA || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || | | Vector DNA || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 0.3 || 0.3 | ||
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| 2x lgn buf (Roche) || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 | | 2x lgn buf (Roche) || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 || 5.0 | ||
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| T4 ligase (NEB) || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || | | T4 ligase (NEB) || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || --- | ||
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| dH<sub>2</sub>O || 2.5 || 2.0 || 1.0 || 3.0 || 2.5 || 2.0 || 1.0 || 3.0 || 3. | | dH<sub>2</sub>O || 2.5 || 2.0 || 1.0 || 3.0 || 2.5 || 2.0 || 1.0 || 3.0 || 3.7 || 4.7 | ||
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| || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL | | || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL || 10 μL | ||
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* Add 30 μL DH5α; 5 min/ ice | * Add 30 μL DH5α; 5 min/ ice | ||
* Plate on 100 μg/mL amp; incubate at 37°C | * Plate on 100 μg/mL amp; incubate at 37°C | ||
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3/21/13<br> | |||
Results for plates 9 and 10 | |||
[[Image:KAH032113_plates1.jpg|300px|Hover name]] | |||
Latest revision as of 22:33, 26 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
03/20/13
Assemblies
3/21/13 Results for plates 9 and 10
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