August/21 August 2009

From 2009.igem.org

(Difference between revisions)

Latest revision as of 08:01, 12 October 2009

transformation

to get new plasmid

⑤(RBS+rhlR+terminator+promoter(reg))

⑦（RBS＋cinR)

⑧（RBS＋luxI)

check the result of transformation

Checked the cell cultures transformed and plated out yesterday (8/10) for colony formation and made an approximate count of the number of colonies.

(plate number)-(location on plate) (no. of colonies)

            [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G]                         4
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_F2621 2-21F]                        0    
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13521 2-6O]                         2
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_E2050 2-13N]                       <15
             ⑤                          0

restriction and digestion

K204009

Vector		Insert
terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])	10	①-3(RBS+LasR)	12
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	4	dH₂O	4
total	20uL	total	20uL

K204010

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

K204011

Vector		Insert
p(tetR)([http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 1-6I])	10	RBS+GYP+terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0240 1-12M])	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	6	dH₂O	4
total	20uL	total	20uL

↓
37°C, 2hr

↓
gel electrophoresis
gel cut

'8/22
purification by [QIAquick Nucleotide Removal Kit]
↓
ligation

ligation
DNA        44
10* buffer 5
ligation   1
total      50uL
↓
16°C , 4hr

back to NOTES

@@ Line 5: / Line 5: @@
 ::⑦（RBS＋cinR)
 ::⑧（RBS＋luxI)
 ===check the result of transformation===
+Checked the cell cultures transformed and plated out yesterday (8/10) for colony formation and made an approximate count of the number of colonies. <br>
+ (plate number)-(location on plate) (no. of colonies)<br>
+             [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G]                         4
+             [http://partsregistry.org/wiki/index.php?title=Part:BBa_F2621 2-21F]                        0
+             [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13521 2-6O]                         2
+             [http://partsregistry.org/wiki/index.php?title=Part:BBa_E2050 2-13N]                       <15
+              ⑤                          0
--8G     4
--21F    0
--6O     2
--13N   <15
- ⑤　　　　　　0
 ===restriction and digestion===
-ⅰ　パーツ⑨
+'''K204009'''
-Insert
+<table border="1" Frame="box">
-H2O　　　　    　4
+<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-No.2　buffer 　2
+<tr><td>terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])</td><td>10</td><td>①-3(RBS+LasR)</td><td>12</td></tr>
-①ー３　      　12
+<tr><td>EcoRI</td><td>1</td><td>EcoRI</td><td>1</td><tr>
-EcoRI     　　1
+<tr><td>XbaI</td><td>1</td><td>SpeI</td><td>1</td><tr>
-SpeⅠ
+<tr><td> No.2</td><td>2</td><td>No.2</td><td>2</td></tr>
+<tr><td>dH<sub>2</sub>O</td><td>4</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>
+'''K204010'''
+<table border="1" Frame="box">
+<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
+<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>10</td><td>EpsE+terminater(②-A)</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 </td><td>2</td><td>No.2</td><td>2</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>
+'''K204011'''
+<table border="1" Frame="box">
+<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
+<tr><td>p(tetR)([http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 1-6I])</td><td>10</td><td>RBS+GYP+terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0240 1-12M])</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 </td><td>2</td><td>No.2</td><td>2</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>
+↓<br>
+°C, 2hr<br>
+↓<br>
+gel electrophoresis<br>
+gel cut<br>
+<br>
+<br>
+'8/22<br>
+purification by [QIAquick Nucleotide Removal Kit]<br>
+↓<br>
+ligation
+ ligation
+ DNA        44
+* buffer 5
+ ligation   1
+ total      50uL
+ ↓
+°C , 4hr
+[https://2009.igem.org/Team:Osaka/NOTES back to NOTES]