User:Karmella Haynes/Notebook/BioBrick cloning/Entry Base: Difference between revisions
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---- | ---- | ||
'''Minipreps'''<br> | '''Minipreps'''<br> | ||
* Check with E/P digests | |||
{| | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
|-valign="top" | |- valign="top" | ||
| | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br> | | rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br> | ||
| rowspan="7" | <!-- [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | | rowspan="7" | <!-- [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | ||
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# BioBrick name: 5' part/(a/b)/size + 3' part/(c/d)/size | # BioBrick name: 5' part/(a/b)/size + 3' part/(c/d)/size | ||
{| | * Digests (Fermentas FD) | ||
|-valign="top" | ** Specific notes | ||
| | |||
| rowspan=" | {| {{table}} cellspacing="3" <!-- Digest rxn. table --> | ||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <!-- [[Image:GelImage.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | |||
|- | |- | ||
| DNA (plasmid) || up to 25 μL | | DNA (plasmid) || up to 25 μL | ||
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* Measure conc.'s | |||
{| | {| {{table}} cellspacing="3" <!-- [DNA] table --> | ||
|- | |- bgcolor=#cfcfcf | ||
| | | Sample || OD260 || 260/280 || ng/μL | ||
|- | |- | ||
| 1. Digested part (a/b) || --- || --- || --- | | 1. Digested part (a/b) || --- || --- || --- | ||
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* Dephosphorylation (Roche) | |||
{| | {| {{table}} cellspacing="3" <!-- Dephos table --> | ||
|- | |- | ||
| | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | |||
|- | |- | ||
| DNA (clean digest) || up to 17 μL (500 ng) | | DNA (clean digest) || up to 17 μL (500 ng) | ||
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* Ligations | |||
{| | {| {{table}} cellspacing="3" <!-- Ligations table --> | ||
| | |- bgcolor=#cfcfcf | ||
| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> mm/dd/yy</font> | |||
|- | |- | ||
| 1. insert(a/b)/size, ## ng + vector(c/d)/size, ## ng || <font color="blue">new BioBrick #:1 (Pick #)</font> | | 1. insert(a/b)/size, ## ng + vector(c/d)/size, ## ng || <font color="blue">new BioBrick #:1 (Pick #)</font> | ||
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|} | |} | ||
{| | {| {{table}} cellspacing="3" <!-- Ligation rxn table --> | ||
| || 1 || 2 || | | || 1 || 2 || | ||
|- | |- |
Revision as of 14:35, 1 December 2012
Karmella's BioBrick Cloning | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
mm/dd/yy
Minipreps
Assemblies
Oligo annealing
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