August/20 August 2009

From 2009.igem.org

(Difference between revisions)

Latest revision as of 08:00, 12 October 2009

1.digestion and ligation

digestion with restriction enzyme

K204007 , (K204003)

Vector		Insert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])	10	cinR([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 1-14F])	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	6	dH₂O	4
total	20uL	total	20uL

K204008

Vector		Insert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])	10	LuxI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0161 2-8O])	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	6	dH₂O	4
total	20uL	total	20uL

↓
37°C, 2hr

↓
gel electrophoresis

UP   : 1-2M , 1-14F , 1-2M , 2-8O
DOWN : No.3-A,B,C,D , No.4-A,B,C,D , No.6-A,B,C,D , Vector

gel cut
↓
purification by [QIAquick Nucleotide Removal Kit]
↓
ligation

ligation
DNA        44
10* buffer 5
ligation   1
total      50uL
↓
16°C overnight

2. rapid check

③(RBS+cinR),K204003

③-C,D : OK

④(RBS+LasR),K204004

④-A,B,C : OK

⑥(RBS+cinI),K204006

⑥-A,B,C : OK

back to NOTES

@@ Line 6: / Line 6: @@
 <table border="1" Frame="box">
 <tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-<tr><td>RBS(1-2M)</td><td>10</td><td>cinR(1-14F)</td><td>12</td></tr>
+<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>10</td><td>cinR([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 1-14F])</td><td>12</td></tr>
 <tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
 <tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
 <tr><td> No.2 Buffer</td><td>2</td><td>No.2</td><td>2</td></tr>
-<tr><td>dH2O</td><td>6</td><td>dH2O</td><td>4</td></tr>
+<tr><td>dH<sub>2</sub>O</td><td>6</td><td>dH<sub>2</sub>O</td><td>4</td></tr>
 <tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
 </table>
@@ Line 18: / Line 18: @@
 <table border="1" Frame="box">
 <tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-<tr><td>RBS(1-2M)</td><td>10</td><td>LuxI(2-8O)</td><td>12</td></tr>
+<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>10</td><td>LuxI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0161 2-8O])</td><td>12</td></tr>
 <tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
 <tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
 <tr><td> No.2 Buffer</td><td>2</td><td>No.2</td><td>2</td></tr>
-<tr><td>dH2O</td><td>6</td><td>dH2O</td><td>4</td></tr>
+<tr><td>dH<sub>2</sub>O</td><td>6</td><td>dH<sub>2</sub>O</td><td>4</td></tr>
 <tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
 </table>
@@ Line 33: / Line 33: @@
 ↓<br>
 gel electrophoresis<br>
+[[Image:Osaka090819.jpg]]<br>
+ UP   : 1-2M , 1-14F , 1-2M , 2-8O
+ DOWN : No.3-A,B,C,D , No.4-A,B,C,D , No.6-A,B,C,D , Vector
 gel cut<br>
 ↓<br>
@@ Line 57: / Line 60: @@
 ⑥(RBS+cinI),K204006<br>
   ⑥-A,B,C : OK
+[https://2009.igem.org/Team:Osaka/NOTES back to NOTES]