User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/10: Difference between revisions
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== | ==01/10/13== | ||
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* | * Violacein: successful growth; re-streak | ||
* | * Golden gate/ transformation: transform DH5α-Turbo with gg assemblies from 1/09/13 | ||
* Golden gate trial 2: new PCR primers | |||
---- | ---- | ||
''' | '''Violacein'''<br> | ||
* | * Plate 1: Kan plasmid | ||
* Plate 2: Amp plasmid (violacein exp.) | |||
---- | ---- | ||
''' | '''Golden gate trial 1''' | ||
* Add total 10.0 μL run to 50 μL DH5α-Turbo; incubate on ice 10 min.; plate on amp agar | |||
---- | |||
* | '''Golden gate trial 2''' | ||
* Re-designed primers to include "handle" preceding the BsmBI sites and a spacer between BsmBI and a 4 bp overlap region | |||
{| {{table}} cellspacing="3" <!-- Digest rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <!-- [[Image:GelImage.jpg| | | rowspan="7" | <u>Expected:</u><br>1. pSB1A3 = ~2000<br>2. hPCD = 186<br>3. BL01 = 2520 | ||
| 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] --> | |||
|- | |- | ||
| | | DNA(plasmid) || 0.2 μL | ||
|- | |- | ||
| | | primer 1 || 1.0 | ||
|- | |- | ||
| | | primer 2 || 1.0 | ||
|- | |- | ||
| | | 2x GoTaq || 25.0 | ||
|- | |- | ||
| | | dH<sub>2</sub>O || 22.8 | ||
|- | |- | ||
| | | || 50.0 μL | ||
|} | |} | ||
PCR | |||
* 95°C, 3 min. | |||
* [95°C, 3 min.; 95°C, 3 min.; 95°C, 3 min.] x35 | |||
* 72°C, 3 min. | |||
* 4°C, ∞ | |||
Revision as of 18:10, 10 January 2013
Karmella's BioBrick Cloning | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||
01/10/13
Violacein
Golden gate trial 1
Golden gate trial 2
PCR
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