Team:Paris/July 19

From 2008.igem.org

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Line 24: Line 24:
===gel 1===
===gel 1===
 +
[[Image:KR000014.jpg|550px|right]]
{| border="1"  
{| border="1"  
 +
|align="center"|'''Band'''
|align="center"|'''Biobricks'''
|align="center"|'''Biobricks'''
|align="center"|'''Clone'''
|align="center"|'''Clone'''
Line 30: Line 32:
|align="center"|'''Measured 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="left"|E1010
|align="center"|1
|align="center"|1
Line 35: Line 44:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|3
|align="left"|E1010
|align="left"|E1010
|align="center"|2
|align="center"|2
Line 40: Line 50:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|4
|align="left"|C0079
|align="left"|C0079
|align="center"|1
|align="center"|1
Line 45: Line 56:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|5
|align="left"|C0079
|align="left"|C0079
|align="center"|2
|align="center"|2
Line 50: Line 62:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|6
|align="left"|C0079
|align="left"|C0079
|align="center"|3
|align="center"|3
Line 55: Line 68:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|7
|align="left"|J23101 : problem !!!
|align="left"|J23101 : problem !!!
|align="center"|1
|align="center"|1
Line 60: Line 74:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|8
|align="left"|J23101 : problem !!!
|align="left"|J23101 : problem !!!
|align="center"|2
|align="center"|2
Line 65: Line 80:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|9
|align="left"|J23101 : problem !!!  
|align="left"|J23101 : problem !!!  
|align="center"|3
|align="center"|3
Line 70: Line 86:
|align="center"|~6-8 kb
|align="center"|~6-8 kb
|-
|-
 +
|align="left"|10
|align="left"|B0034
|align="left"|B0034
|align="center"|2
|align="center"|2
Line 75: Line 92:
|align="center"|~2 kb
|align="center"|~2 kb
|-
|-
 +
|align="left"|11
|align="left"|B0034
|align="left"|B0034
|align="center"|X
|align="center"|X
Line 80: Line 98:
|align="center"|~2 kb
|align="center"|~2 kb
|-
|-
 +
|align="left"|12
|align="left"|B0034 : problem !!!
|align="left"|B0034 : problem !!!
|align="center"|Y
|align="center"|Y
Line 85: Line 104:
|align="center"|~5 kb
|align="center"|~5 kb
|-
|-
 +
|align="left"|13
|align="left"|E0040
|align="left"|E0040
|align="center"|1
|align="center"|1
Line 90: Line 110:
|align="center"|~2.5-3 kb
|align="center"|~2.5-3 kb
|-
|-
 +
|align="left"|14
|align="left"|E0040
|align="left"|E0040
|align="center"|2
|align="center"|2
Line 95: Line 116:
|align="center"|~2.5-3 kb
|align="center"|~2.5-3 kb
|-
|-
 +
|align="left"|15
|align="left"|R0040
|align="left"|R0040
|align="center"|?
|align="center"|?
Line 100: Line 122:
|align="center"|~2 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!
===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!
Line 109: Line 238:
===PCR===
===PCR===
For 1 tube :  
For 1 tube :  
 +
25µL Quick Load<br>
25µL Quick Load<br>
1µL Oligo F<br>
1µL Oligo F<br>
Line 118: Line 248:
===Electrophoresis===
===Electrophoresis===
-
Band 0 : ladder 100pb
+
[[Image:screen.jpg|right|250px]]
-
Band 2 : B0034 clone 1 -> no result
+
{| border="1"
-
Band 3 : B0034 clone 2 -> ok
+
|align="center"|'''Band'''
-
Band 4 : R0040 clone 1 -> ok
+
|align="center"|'''Biobricks'''
-
Band 5 : R0040 clone 2 -> problem
+
|align="center"|'''Clone'''
-
Band 6 : R0040 ? -> problem
+
|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"