User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/10: Difference between revisions
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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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| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1. pSB1A3 = ~2000<br>2. hPCD = 186<br>3. BL01 = 2520 | | rowspan="7" | <u>Expected:</u><br>1. pSB1A3 = ~2000<br>2. hPCD = 186<br>3. BL01 = 2520 | ||
| rowspan="7" | [[Image: | | rowspan="7" | [[Image:KAH011013_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || 0.2 μL | | DNA(plasmid) || 0.2 μL | ||
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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, ∞ | |||
Revision as of 21:42, 18 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
Thermal cycling
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