August/8 October 2009

From 2009.igem.org

(Difference between revisions)

Latest revision as of 01:31, 21 October 2009

1.sequence check
Today we conducted PCR in order to sequence the parts we have built over the summer. Using the chain-termination method and standard sequencing primers from the Parts Registry, we obtained chain-terminated clones of each sample and put them through an automatic DNA sequencer. The results will be picked up and analyzed tomorrow.

2.colony check

sample        no. of colony
 2-6O                +++
  11                   +++ 
  20                   ++
  21                   +++
 1-15L               +++
  1-15J              +++
   52                    20
   67                     2
   68                     0
   69                     0

3.ligation

sample 
 70 , 71 , 74 , 75

4.digation

K204059

Vector		Insert
1-8E	6	1-8K	1
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	8	dH₂O	13
total	20uL	total	20uL

K204072

Vector		Insert
2-8E	2	31	6
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	8
total	20uL	total	20uL

↓
37 degree , 3hr
gel cut & ligation

5.transformation

sample 
70 , 71 , 74 , 75 , X4 , X4

6.color intensity check

plac+color , ptet+color

7.sequence check

sample 
22 , 47 , 40 , 25 , 51 , 50 , 31 , 32

back to NOTES

@@ Line 1: / Line 1: @@
-Today we conducted PCR in order to sequence the parts we have built over the summer. Using the chain-termination method and standard sequencing primers from the Parts Registry, we obtained chain-terminated clones of each sample and put them through an automatic DNA sequencer. The results will be picked up and analyzed tomorrow.
+.sequence check<br>
+Today we conducted PCR in order to sequence the parts we have built over the summer. Using the chain-termination method and standard sequencing primers from the Parts Registry, we obtained chain-terminated clones of each sample and put them through an automatic DNA sequencer. The results will be picked up and analyzed tomorrow.<br>
+.colony check<br>
+ sample        no. of colony
+-6O                +++
+                   +++
+                   ++
+                   +++
+-15L               +++
+-15J              +++
+                    20
+                     2
+                     0
+                     0
+.ligation <br>
+ sample
+, 71 , 74 , 75
+.digation<br>
+K204059
+<table border="1" Frame="box">
+<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
+<tr><td>1-8E</td><td>6</td><td>1-8K</td><td>1</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>8</td><td>dH<sub>2</sub>O</td><td>13</td></tr>
+<tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
+</table>
+K204072
+<table border="1" Frame="box">
+<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
+<tr><td>2-8E</td><td>2</td><td>31</td><td>6</td></tr>
+<tr><td>EcoRI</td><td>1</td><td>EcoRI</td><td>1</td><tr>
+<tr><td>XbaI</td><td>1</td><td>SpeI</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>12</td><td>dH<sub>2</sub>O</td><td>8</td></tr>
+<tr><td>total</td><td>20uL</td><td>total</td><td>20uL</td></tr>
+</table>
+<br>
+↓<br>
+degree , 3hr<br>
+gel cut & ligation<br>
+.transformation<br>
+ sample
+, 71 , 74 , 75 , X4 , X4
+<br>
+.color intensity check<br>
+ plac+color , ptet+color
+.sequence check<br>
+ sample
+, 47 , 40 , 25 , 51 , 50 , 31 , 32
+[https://2009.igem.org/Team:Osaka/NOTES back to NOTES]