Team:Paris/July 19

From 2008.igem.org

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{{Paris/Calendar_Links|July 18|July 20}}
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== MiniPrep==
== MiniPrep==
Line 20: Line 23:
== Quantitative analysis of ADN ==
== Quantitative analysis of ADN ==
-
photo gel 1
+
===gel 1===
 +
[[Image:KR000014.jpg|550px|right]]
 +
{| border="1"
 +
|align="center"|'''Band'''
 +
|align="center"|'''Biobricks'''
 +
|align="center"|'''Clone'''
 +
|align="center"|'''Expected size'''
 +
|align="center"|'''Measured size'''
 +
|-
 +
|align="left"|1
 +
|align="left"|Ladder 1 kpb
 +
|align="center"| 
 +
|align="center"| 
 +
|align="center"| 
 +
|-
 +
|align="left"|2
 +
|align="left"|E1010
 +
|align="center"|1
 +
|align="center"|5.106 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|3
 +
|align="left"|E1010
 +
|align="center"|2
 +
|align="center"|5.106 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|4
 +
|align="left"|C0079
 +
|align="center"|1
 +
|align="center"|5.181 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|5
 +
|align="left"|C0079
 +
|align="center"|2
 +
|align="center"|5.181 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|6
 +
|align="left"|C0079
 +
|align="center"|3
 +
|align="center"|5.181 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|7
 +
|align="left"|J23101 : problem !!!
 +
|align="center"|1
 +
|align="center"|3.037 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|8
 +
|align="left"|J23101 : problem !!!
 +
|align="center"|2
 +
|align="center"|3.037 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|9
 +
|align="left"|J23101 : problem !!!
 +
|align="center"|3
 +
|align="center"|3.037 kb
 +
|align="center"|~6-8 kb
 +
|-
 +
|align="left"|10
 +
|align="left"|B0034
 +
|align="center"|2
 +
|align="center"|2.091 kb
 +
|align="center"|~2 kb
 +
|-
 +
|align="left"|11
 +
|align="left"|B0034
 +
|align="center"|X
 +
|align="center"|2.091 kb
 +
|align="center"|~2 kb
 +
|-
 +
|align="left"|12
 +
|align="left"|B0034 : problem !!!
 +
|align="center"|Y
 +
|align="center"|2.091 kb
 +
|align="center"|~5 kb
 +
|-
 +
|align="left"|13
 +
|align="left"|E0040
 +
|align="center"|1
 +
|align="center"|2.799 kb
 +
|align="center"|~2.5-3 kb
 +
|-
 +
|align="left"|14
 +
|align="left"|E0040
 +
|align="center"|2
 +
|align="center"|2.799 kb
 +
|align="center"|~2.5-3 kb
 +
|-
 +
|align="left"|15
 +
|align="left"|R0040
 +
|align="center"|?
 +
|align="center"|2.133 kb
 +
|align="center"|~2 kb
 +
|}
 +
 
 +
 
We see bad results for B0034 (bands 10 to 12), R0040 (band 15). We'll try again!
We see bad results for B0034 (bands 10 to 12), R0040 (band 15). We'll try again!
-
photo gel 2
+
===gel 2===
 +
[[Image:KR000015.jpg|550px|right]]
 +
{| border="1"
 +
|align="center"|'''Band'''
 +
|align="center"|'''Biobricks'''
 +
|align="center"|'''Clone'''
 +
|align="center"|'''Expected size'''
 +
|align="center"|'''Measured size'''
 +
|-
 +
|align="left"|1
 +
|align="left"|Ladder 1 kpb
 +
|align="center"| 
 +
|align="center"| 
 +
|align="center"| 
 +
|-
 +
|align="left"|2
 +
|align="left"|J23109
 +
|align="center"|1
 +
|align="center"|3.037 kb
 +
|align="center"|~ 2-3 kb
 +
|-
 +
|align="left"|3
 +
|align="left"|J23109 : Problem !!!
 +
|align="center"|2
 +
|align="center"|3.037 kb
 +
|align="center"|~ 6-8 kb
 +
|-
 +
|align="left"|4
 +
|align="left"|J23109 : Problem !!!
 +
|align="center"|3
 +
|align="center"|3.037 kb
 +
|align="center"|> 10 kb
 +
|-
 +
|align="left"|5
 +
|align="left"|pSB1A3
 +
|align="center"|1
 +
|align="center"|2.157 kb
 +
|align="center"|~ 2-3 kb
 +
|-
 +
|align="left"|6
 +
|align="left"|pSB1A3
 +
|align="center"|2
 +
|align="center"|2.157 kb
 +
|align="center"|~ 2-3 kb
 +
|-
 +
|align="left"|7
 +
|align="left"|pSB1A3
 +
|align="center"|3
 +
|align="center"|2.157 kb
 +
|align="center"|~ 2-3 kb
 +
|-
 +
|align="left"|8
 +
|align="left"|E0030
 +
|align="center"|1
 +
|align="center"|3.912 kb
 +
|align="center"|~ 3.5 kb
 +
|-
 +
|align="left"|9
 +
|align="left"|E0030
 +
|align="center"|2
 +
|align="center"|3.912 kb
 +
|align="center"|~ 3.5 kb
 +
|-
 +
|align="left"|10
 +
|align="left"|E0030
 +
|align="center"|3
 +
|align="center"|3.912 kb
 +
|align="center"|~ 3.5 kb
 +
|-
 +
|align="left"|11
 +
|align="left"|R0079
 +
|align="center"|3
 +
|align="center"|2.236 kb
 +
|align="center"|~ 1.5 kb
 +
|-
 +
|align="left"|12
 +
|align="left"|R0079
 +
|align="center"|4
 +
|align="center"|2.236 kb
 +
|align="center"|~ 1.5 kb
 +
|-
 +
|align="left"|13
 +
|align="left"|R0079
 +
|align="center"|6
 +
|align="center"|2.236 kb
 +
|align="center"|~ 1.5 kb
 +
|-
 +
|align="left"|14
 +
|align="left"|C0179 : Problem !!!
 +
|align="center"|1
 +
|align="center"|2.802 kb
 +
|align="center"|> 10 kb
 +
|-
 +
|align="left"|15
 +
|align="left"|C0179 : Problem !!!
 +
|align="center"|2
 +
|align="center"|2.802 kb
 +
|align="center"|~ 10 kb
 +
|-
 +
|align="left"|16
 +
|align="left"|C0179
 +
|align="center"|3
 +
|align="center"|2.802 kb
 +
|align="center"|~ 2.5 kb
 +
|}
We see bad results for J23109 (bands 1 to 3), C0179 (bands 14 to 16). We'll try again!
We see bad results for J23109 (bands 1 to 3), C0179 (bands 14 to 16). We'll try again!
-
 
== Screening R0040 and B0034 ==
== Screening R0040 and B0034 ==
===PCR===
===PCR===
For 1 tube :  
For 1 tube :  
 +
25µL Quick Load<br>
25µL Quick Load<br>
1µL Oligo F<br>
1µL Oligo F<br>
1µL Oligo R<br>
1µL Oligo R<br>
-
23µL H<math>_2</math>O
+
23µL H<sub>2</sub>O
 +
 
 +
Program SCREENIN
 +
 
 +
===Electrophoresis===
 +
 
 +
[[Image:screen.jpg|right|250px]]
 +
{| border="1"
 +
|align="center"|'''Band'''
 +
|align="center"|'''Biobricks'''
 +
|align="center"|'''Clone'''
 +
|align="center"|'''Expected size'''
 +
|align="center"|'''Measured size'''
 +
|-
 +
|align="left"|2
 +
|align="left"|Ladder 100 pb
 +
|align="center"|&nbsp;
 +
|align="center"|&nbsp;
 +
|align="center"|&nbsp;
 +
|-
 +
|align="left"|3
 +
|align="left"|B0034
 +
|align="center"|1
 +
|align="center"|250 pb
 +
|align="center"|nothing
 +
|-
 +
|align="left"|4
 +
|align="left"|B0034
 +
|align="center"|2
 +
|align="center"|250 pb
 +
|align="center"|~250pb
 +
|-
 +
|align="left"|5
 +
|align="left"|R0040
 +
|align="center"|1
 +
|align="center"|292 pb
 +
|align="center"|~300pb
 +
|-
 +
|align="left"|6
 +
|align="left"|R0040 : Problem !!!
 +
|align="center"|2
 +
|align="center"|292 pb
 +
|align="center"|~250pb
 +
|-
 +
|align="left"|7
 +
|align="left"|R0040 : Problem !!!
 +
|align="center"|?
 +
|align="center"|292 pb
 +
|align="center"|~250pb
 +
|-
 +
|}

Latest revision as of 12:54, 22 July 2008

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Contents

MiniPrep

The protocol of MiniPrep of Quiagen has been done on all the clones cultivated on the 18th. J23109 clones 1,2,5
E0030 clones 1,2,3
pSB1A3 clones 1,2,3
C0179 clones 1,2,7
C0079 clones 1,2,3
R0079 clones 3,4,6
E1010 clones 1,2,3
B0034 clones 1,2,3
J23101 clones 1,2,3
E0040 clones 1,2
R0040 clones 1,2


We did an error during the manipulation (probably an exchange between some tubes) so we decided to screen again clones of B0034 and R0040.


Quantitative analysis of ADN

gel 1

KR000014.jpg
Band Biobricks Clone Expected size Measured size
1 Ladder 1 kpb      
2 E1010 1 5.106 kb ~6-8 kb
3 E1010 2 5.106 kb ~6-8 kb
4 C0079 1 5.181 kb ~6-8 kb
5 C0079 2 5.181 kb ~6-8 kb
6 C0079 3 5.181 kb ~6-8 kb
7 J23101 : problem !!! 1 3.037 kb ~6-8 kb
8 J23101 : problem !!! 2 3.037 kb ~6-8 kb
9 J23101 : problem !!! 3 3.037 kb ~6-8 kb
10 B0034 2 2.091 kb ~2 kb
11 B0034 X 2.091 kb ~2 kb
12 B0034 : problem !!! Y 2.091 kb ~5 kb
13 E0040 1 2.799 kb ~2.5-3 kb
14 E0040 2 2.799 kb ~2.5-3 kb
15 R0040 ? 2.133 kb ~2 kb


We see bad results for B0034 (bands 10 to 12), R0040 (band 15). We'll try again!

gel 2

KR000015.jpg
Band Biobricks Clone Expected size Measured size
1 Ladder 1 kpb      
2 J23109 1 3.037 kb ~ 2-3 kb
3 J23109 : Problem !!! 2 3.037 kb ~ 6-8 kb
4 J23109 : Problem !!! 3 3.037 kb > 10 kb
5 pSB1A3 1 2.157 kb ~ 2-3 kb
6 pSB1A3 2 2.157 kb ~ 2-3 kb
7 pSB1A3 3 2.157 kb ~ 2-3 kb
8 E0030 1 3.912 kb ~ 3.5 kb
9 E0030 2 3.912 kb ~ 3.5 kb
10 E0030 3 3.912 kb ~ 3.5 kb
11 R0079 3 2.236 kb ~ 1.5 kb
12 R0079 4 2.236 kb ~ 1.5 kb
13 R0079 6 2.236 kb ~ 1.5 kb
14 C0179 : Problem !!! 1 2.802 kb > 10 kb
15 C0179 : Problem !!! 2 2.802 kb ~ 10 kb
16 C0179 3 2.802 kb ~ 2.5 kb

We see bad results for J23109 (bands 1 to 3), C0179 (bands 14 to 16). We'll try again!

Screening R0040 and B0034

PCR

For 1 tube :

25µL Quick Load
1µL Oligo F
1µL Oligo R
23µL H2O

Program SCREENIN

Electrophoresis

Screen.jpg
Band Biobricks Clone Expected size Measured size
2 Ladder 100 pb      
3 B0034 1 250 pb nothing
4 B0034 2 250 pb ~250pb
5 R0040 1 292 pb ~300pb
6 R0040 : Problem !!! 2 292 pb ~250pb
7 R0040 : Problem !!! ? 292 pb ~250pb
Retrieved from "http://2008.igem.org/Team:Paris/July_19"