Difference between revisions of "Part:BBa K1429002"

Revision as of 01:35, 1 October 2014

Cyan Fluorescent Protein (CFP) "Cindy Lou" coding region, intellectual property-free

IP free cyan fluorescent protein. We have a ribosome binding site (RBS).

Component	Using purified digest product	Using unpurified digest product	BB alone
dH2O	x	11	14
Insert (RFP or GFP)	14	3	x
1:10 BB	3	3	3
T4 buffer (10	1	1	1

Sequence and Features

Assembly Compatibility:

@@ Line 4: / Line 4: @@
 IP free cyan fluorescent protein.  We have a ribosome binding site (RBS).
-Background
-Making sure the inserts are here from the colony PCR results - are the GFP and RFP inserts present?
+<table style="width: 500px;">
+	<tbody>
+		<tr>
+			<td><strong>Component</strong></td>
+			<td><strong>Using purified digest product</strong></td>
+			<td><strong>Using unpurified digest product</strong></td>
+			<td><strong>BB alone</strong></td>
+		</tr>
+		<tr>
+			<td>dH2O</td>
+			<td>x</td>
+			<td>11</td>
+			<td>14</td>
+		</tr>
+		<tr>
+			<td>Insert (RFP or GFP)</td>
+			<td>14</td>
+			<td>3</td>
+			<td>x</td>
+		</tr>
+		<tr>
+			<td>1:10 BB</td>
+			<td>3</td>
+			<td>3</td>
+			<td>3</td>
+		</tr>
+		<tr>
+			<td>T4 buffer (10</td>
+			<td>1</td>
+			<td>1</td>
+			<td>1</td>
+		</tr>
+		<tr>
+			<td>&nbsp;</td>
+			<td>&nbsp;</td>
+			<td>&nbsp;</td>
+			<td>&nbsp;</td>
+		</tr>
+	</tbody>
+</table>
-Digest each plasmid with EcoRI/PstI. Incubate 15' at 37C.
-Run entire digest on 0.8% gel.
-Results
-These were slightly confounding. What I *think* happened is that the buffer wasn't optimal for one of the enzymes, so the digest wasn't run to completion. Why do I think this?
 <!-- Add more about the biology of this part here