User:Karmella Haynes/Notebook/BioBrick cloning/2015/04/28: Difference between revisions
From OpenWetWare
Line 33: | Line 33: | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1-4. Bra/MRV = ~3200, 776<br>Empty MRV E/X = ~3200<br>5. pAub/AubR/MRV = 4240<br>6. pBja/BjaR/MRV = 4099<br>7. pRpa/RpaR/MRV = 4116<br>Empty vector = ~4000, 971 | | rowspan="7" | <u>Expected:</u><br>1-4. Bra/MRV = ~3200, 776<br>Empty MRV E/X = ~3200<br>5. pAub/AubR/MRV = 4240<br>6. pBja/BjaR/MRV = 4099<br>7. pRpa/RpaR/MRV = 4116<br>Empty vector = ~4000, 971 | ||
| rowspan="7" | | | rowspan="7" | [[Image:KAH042815_gel1.jpg|250px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || 3.0 μL | | DNA(plasmid) || 3.0 μL | ||
Line 111: | Line 111: | ||
| bgcolor=#cfcfcf | Mix (x2) | | bgcolor=#cfcfcf | Mix (x2) | ||
| rowspan=7 | Expected:<br>1. pAubR = 153<br>2. pBjaR = 122<br>3. pBraR = 214<br>4. pRpaR = 136 | | rowspan=7 | Expected:<br>1. pAubR = 153<br>2. pBjaR = 122<br>3. pBraR = 214<br>4. pRpaR = 136 | ||
| rowspan=7 | [[Image:KAH042815_gel2.jpg|250px|Hover name]]<br>5 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |- | ||
| gBlock DNA (2 ng/μL) || 0.5 || 1.0 | | gBlock DNA (2 ng/μL) || 0.5 || 1.0 | ||
Line 142: | Line 143: | ||
| Sample || OD260 || 260/280 || ng/μL | | Sample || OD260 || 260/280 || ng/μL | ||
|- | |- | ||
| 1. pAubR PCR || | | 1. pAubR PCR || ### || ### || ### | ||
|- | |- | ||
| 2. pBjaR PCR || 0.177 || 1.904 || 177.3 | | 2. pBjaR PCR || 0.177 || 1.904 || 177.3 |
Revision as of 18:42, 28 April 2015
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> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
04/28/15
Minipreps
Ryan - PCR & Dig/Lig (promoter insert trial #2) Ryan - Receiver plasmids, assembly - REVISED STRATEGY (PCR promoters)
Thermal cycler: Labnet - GOTAQ
--> 37°C/ 10 min.; 75°C/ 2 min.; [final] = ~1 ng/μL
Thermal cycler: Labnet - BbsI Dig/Lig
|