August/20 August 2009

From 2009.igem.org

(Difference between revisions)
(New page: 1.<B>digestion and ligation</b><br> digestion with restriction enzyme<br> '''K204007''' , (K204003) <table border="1" Frame="box"> <tr><th colspan="2">Vector</th><th colspan="2">Insert</...)
Line 10: Line 10:
<tr><td>PstI</td><td>1</td><td>PstI</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> 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>
<tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
</table>
</table>
Line 22: Line 22:
<tr><td>PstI</td><td>1</td><td>PstI</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> 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>
<tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
</table>
</table>

Revision as of 06:29, 23 August 2009

1.digestion and ligation

digestion with restriction enzyme

K204007 , (K204003)

VectorInsert
RBS(1-2M)10cinR(1-14F)12
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O6dH2O4
total20uLtotal20uL


K204008

VectorInsert
RBS(1-2M)10LuxI(2-8O)12
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O6dH2O4
total20uLtotal20uL


↓
37°C, 2hr

↓
gel electrophoresis
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
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