Team:TU Delft/23 July 2010 content

From 2010.igem.org

(Difference between revisions)
(→Alkane Degradation)
(→Alkane Degradation)
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With a number of [https://2010.igem.org/Team:TU_Delft#/blog?blog=22_July_2010 yesterday's] colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description.
With a number of [https://2010.igem.org/Team:TU_Delft#/blog?blog=22_July_2010 yesterday's] colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description.
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
|'''#'''
|'''#'''
|'''Contents'''
|'''Contents'''
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|'''Buffer'''
|'''Buffer'''
|'''BSA'''
|'''BSA'''
-
|'''Expected size(s)'''
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|'''Expected size(s) (bp)'''
|-
|-
|1
|1
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
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|3 (Biolabs)
|no
|no
 +
|1341, 2455
|-
|-
|3
|3
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
+
|3 (Biolabs)
|no
|no
 +
|276, 2455
|-
|-
|4
|4
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
+
|3 (Biolabs)
|no
|no
 +
|285, 2455
|-
|-
|5
|5
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
+
|3 (Biolabs)
|no
|no
 +
|285, 2455
|-
|-
|6
|6
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
+
|3 (Biolabs)
|no
|no
 +
|1382, 2455
|-
|-
|7
|7
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|0.5 μl EcoRI
|0.5 μl EcoRI
|0.5 μl PstI
|0.5 μl PstI
-
|Buffer 3 (Biolabs)
+
|3 (Biolabs)
|no
|no
 +
|855, 2455
 +
|-
 +
|8
 +
|Empty
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|-
 +
|9
 +
|007T (2)
 +
|1 μl PstI
 +
|1 μl PvuII
 +
|H (Boehringer)
 +
|no
 +
|2529,983,251
 +
|-
 +
|10
 +
|008T (4)
 +
|1 μl AatII
 +
|0.5 μl KpnI
 +
|A (Boehringer)
 +
|1 μl
 +
|271, 2427
 +
|-
 +
|11
 +
|009T (6)
 +
|0.5 μl KpnI
 +
|1 μl NruI
 +
|B (Boehringer)
 +
|1 μl
 +
|598, 2109
 +
|-
 +
|12
 +
|009T (7)
 +
|0.5 μl KpnI
 +
|1 μl NruI
 +
|B (Boehringer)
 +
|1 μl
 +
|598, 2109
 +
|-
 +
|13
 +
|010T (9)
 +
|1 μl EarI
 +
|0.5 μl PstI
 +
|1 (Biolabs)
 +
|no
 +
|1075, 2729
 +
|-
 +
|14
 +
|017T (14)
 +
|0.5 μl AdhI
 +
|0.5 μl SpeI
 +
|4 (Biolabs)
 +
|no
 +
|428, 2849
 +
|-
 +
|15
 +
|EZ load (ladder)
 +
|
 +
|
 +
|
 +
|
 +
|n/a
|-
|-
-
 
-
 
|}
|}
-
 
+
[[Image:TUDelft_Digestion_Kira_23-07.png|500px|thumb|left|Digestion check]]
-
 
+
-
 
+
-
[[Image:TUDelft_Digestion_Kira_23-07.png|200px|thumb|left|Digestion check]]
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Revision as of 10:53, 26 July 2010

Alkane Degradation

With a number of yesterday's colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description.

# Contents Enzyme 1 Enzyme 2 Buffer BSA Expected size(s) (bp)
1 Smartladder (5μl) n/a
2 007T (2) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 1341, 2455
3 008T (4) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 276, 2455
4 009T (6) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 285, 2455
5 009T (7) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 285, 2455
6 010T (9) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 1382, 2455
7 017T (14) 0.5 μl EcoRI 0.5 μl PstI 3 (Biolabs) no 855, 2455
8 Empty
9 007T (2) 1 μl PstI 1 μl PvuII H (Boehringer) no 2529,983,251
10 008T (4) 1 μl AatII 0.5 μl KpnI A (Boehringer) 1 μl 271, 2427
11 009T (6) 0.5 μl KpnI 1 μl NruI B (Boehringer) 1 μl 598, 2109
12 009T (7) 0.5 μl KpnI 1 μl NruI B (Boehringer) 1 μl 598, 2109
13 010T (9) 1 μl EarI 0.5 μl PstI 1 (Biolabs) no 1075, 2729
14 017T (14) 0.5 μl AdhI 0.5 μl SpeI 4 (Biolabs) no 428, 2849
15 EZ load (ladder) n/a
Digestion check
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