August/21 August 2009

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Revision as of 05:44, 22 August 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)

            2-8G                         4
            2-21F                        0    
            2-6O                         2
            2-13N                       <15
             ⑤                          0

restriction and digestion

K204009

Vector		Insert
terminater(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(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)(1-6I)	10	RBS+GYP+terminater(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

@@ 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>
+-8G                         4
+-21F                        0
+-6O                         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(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>dH2O</td><td>4</td><td>dH2O</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(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>dH2O</td><td>6</td><td>dH2O</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)(1-6I)</td><td>10</td><td>RBS+GYP+terminater(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>dH2O</td><td>6</td><td>dH2O</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>