Team:Lethbridge/Notebook/Lab Work/June

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Revision as of 23:00, 10 June 2010

June 2010

June 1/2010

JV quantified the amount of DNA in gels run to date using ImageJ software. Results to be posted in working plasmids box.

June 2/2010

(In Lab: JV)

Objective: Isolate plasmid DNA (BBa_J33204) from DH5α cells and confirm results.

Method: "Mini-prep" the plasmid DNA using boiling lysis miniprep. Then restrict the DNA once and run on a 1% agarose gel (TAE).

Restriction Reaction

Ingredient	Volume(µL)
MilliQ H₂0 Water	15.75
Orange Buffer (10x)	2
pDNA (rbs-xylE)	2
EcoRI	0.25

Unrestricted Control

Ingredient	Volume(µL)
MilliQ H₂0 Water	16
Orange Buffer (10x)	2
pDNA (rbs-xylE)	2

June 3/2010

Objective: Ligate pLacI to sRBS-Lum-dT using the three antibiotic assembly method (according to Tom Knight's protocol). Previous ligation had miniscule quantities of DNA in ligation, not surprising it didn't work.

@@ Line 50: / Line 50: @@
 <b>Method:</b> "Mini-prep" the plasmid DNA using [[Team:Lethbridge/Notebook/Protocols|boiling lysis miniprep]]. Then restrict the DNA once and run on a 1% agarose gel (TAE). <br>
+Restriction Reaction
+<table><table border ="3">
+<tr><td><b>Ingredient</b></td><td>Volume(&micro;L)</td></tr>
+<tr><td>MilliQ H<sub>2</sub>0 Water</td><td>15.75</td></tr>
+<tr><td>Orange Buffer (10x)</td><td>2</td></tr>
+<tr><td>pDNA (rbs-xylE)</td><td>2</td></tr>
+<tr><td>EcoRI</td><td>0.25</td></tr>
+</table>
+Unrestricted Control
+<table><table border ="3">
+<tr><td><b>Ingredient</b></td><td>Volume(&micro;L)</td></tr>
+<tr><td>MilliQ H<sub>2</sub>0 Water</td><td>16</td></tr>
+<tr><td>Orange Buffer (10x)</td><td>2</td></tr>
+<tr><td>pDNA (rbs-xylE)</td><td>2</td></tr>
+</table>
 ===June 3/2010===
 <b>Objective:</b> Ligate pLacI to sRBS-Lum-dT using the three antibiotic assembly method (according to Tom Knight's protocol). Previous ligation had miniscule quantities of DNA in ligation, not surprising it didn't work.<br>