User:University of Washington/13 August 2008

(Difference between revisions)

Revision as of 22:00, 14 August 2008

experiment from Aug. 7 results

experiment #	Bacteria	Yeast	Yeast count	transconjugation efficiency
1	250uL	250uL	66	2.64E-5
2	200uL	300uL	64	2.13E-5
3	100uL	400uL	32	8.00e-6
4	50uL	450uL	18	4.00e-6

- Overnight QuikChange#4 culture from Amp plate (reaction1 x 4, reaction2 x 2) in Tsy+Amp

- Sequence of DNA from QuikChange#3 came back ==> No mutation.

-Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3)

	dH2O(ul)	Buffer(ul)	BSA(ul)	DNA(ul)	E(ul)	X(ul)	X(ul)	S(ul)
BBa_Elowitz	32.5	EcoRI, 5	0.5	10	1	-	1	-
LuxR	32.5	NEB3, 5	0.5	10	-	1	-	1
GFP	32.5	NEB3, 5	0.5	10	-	1	-	1
P1010	32.5	EcoRI, 5	0.5	10	1	-	-	1

@@ Line 8: / Line 8: @@
 <tr><td>3</td><td>100uL</td><td>400uL</td><td>32</td><td>8.00e-6</td></tr>
 <tr><td>4</td><td>50uL</td><td>450uL</td><td>18</td><td>4.00e-6</td></tr>
+</table>
+==LuxR from AraC and TetR(Faifan)==
+<h4>QuikChange Approach</h4>
+- Overnight QuikChange#4 culture from Amp plate (reaction1 x 4, reaction2 x 2) in Tsy+Amp
+- Sequence of DNA from QuikChange#3 came back ==> No mutation.
+<h4>Elowitz's</h4>
+-Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3)
 <table>
+<tr><th></th><th>dH2O(ul)</th><th>Buffer(ul)</th><th>BSA(ul)</th><th>DNA(ul)</th><th>E(ul)</th><th>X(ul)</th><th>X(ul)</th><th>S(ul)</th><th>P(ul)</th></tr>
+<tr><th>BBa_Elowitz</th><th>32.5</th><th>EcoRI, 5</th><th>0.5</th><th>10</th><th>1</th><th>-</th><th>1</th><th>-</th></tr>
+<tr><th>LuxR</th><th>32.5</th><th>NEB3, 5</th><th>0.5</th><th>10</th><th>-</th><th>1</th><th>-</th><th>1</th></tr>
+<tr><th>GFP</th><th>32.5</th><th>NEB3, 5</th><th>0.5</th><th>10</th><th>-</th><th>1</th><th>-</th><th>1</th></tr>
+<tr><th>P1010</th><th>32.5</th><th>EcoRI, 5</th><th>0.5</th><th>10</th><th>1</th><th>-</th><th>-</th><th>1</th></tr>
+</table>
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