Team:TU Delft/28 July 2010 content

From 2010.igem.org

(Difference between revisions)
(→Alkane degradation)
Line 3: Line 3:
The plates containing [https://2010.igem.org/Team:TU_Delft#/blog?blog=27_July_2010 yesterday's] ligations contained colonies, to check whether they really contain the desired BioBrick a [[Team:TU_Delft/protocols/colony PCR|colony PCR]] was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.
The plates containing [https://2010.igem.org/Team:TU_Delft#/blog?blog=27_July_2010 yesterday's] ligations contained colonies, to check whether they really contain the desired BioBrick a [[Team:TU_Delft/protocols/colony PCR|colony PCR]] was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.
-
[[Image:TUDelft_20100728_scPCR1.png|400px|thumb|left|sc PCR part 1]]
+
[[Image:TUDelft_20100728_scPCR1.png|400px|thumb|left|1% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]]
'''Lane Description part 1'''
'''Lane Description part 1'''
Line 9: Line 9:
|'''#'''
|'''#'''
|'''Description'''
|'''Description'''
-
|'''Expected (bp)'''
+
|'''Expected lenght (bp)'''
-
|'''As expected?'''
+
|'''Status'''
 +
|'''Remarks'''
|-
|-
|1
|1
-
|SmartLadder (5 μL)
+
|SmartLadder
 +
|n/a
 +
|n/a
|
|
 +
|-
 +
|2
 +
|J61100 + rubA3
 +
|473
 +
|<font color=limegreen>✓</font>
|
|
|-
|-
-
|2-6
+
|3
|J61100 + rubA3
|J61100 + rubA3
|473
|473
-
|2, 3, 5  
+
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|4
 +
|J61100 + rubA3
 +
|473
 +
|<font color=red>✗</font>
 +
|
 +
|-
 +
|5
 +
|J61100 + rubA3
 +
|473
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|6
 +
|J61100 + rubA3
 +
|473
 +
|<font color=red>✗</font>
 +
|
|-
|-
-
|7-11
+
|7
|J61100 + rubA4
|J61100 + rubA4
|482
|482
-
|7, 8, 9, 10, 11
+
|<font color=limegreen>✓</font>
 +
|
|-
|-
-
|12-16
+
|8
 +
|J61100 + rubA4
 +
|482
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|9
 +
|J61100 + rubA4
 +
|482
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|10
 +
|J61100 + rubA4
 +
|482
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|11
 +
|J61100 + rubA4
 +
|482
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|12
|J61100 + rubR
|J61100 + rubR
|1505
|1505
-
|12, 13, 14, 15, 16
+
|<font color=limegreen>✓</font>
 +
|
|-
|-
-
|17
+
|13
-
|SmartLadder (5 μL)
+
|J61100 + rubR
 +
|1505
 +
|<font color=limegreen>✓</font>
|
|
 +
|-
 +
|14
 +
|J61100 + rubR
 +
|1505
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|15
 +
|J61100 + rubR
 +
|1505
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|16
 +
|J61100 + rubR
 +
|1505
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|17
 +
|SmartLadder
 +
|n/a
 +
|n/a
|
|
|-
|-
|}
|}
-
[[Image:TUDelft_20100728_scPCR2b.png|400px|thumb|left|sc PCR part 2]]
+
[[Image:TUDelft_20100728_scPCR2b.png|400px|thumb|left|% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker]]
'''Lane Description part 2'''
'''Lane Description part 2'''
Line 45: Line 123:
|'''#'''
|'''#'''
|'''Description'''
|'''Description'''
-
|'''Expected (bp)'''
+
|'''Expected lenght (bp)'''
-
|'''As expected?'''
+
|'''Status'''
 +
|'''Remarks'''
|-
|-
|1
|1
-
|SmartLadder (5 μL)
+
|SmartLadder
 +
|n/a
 +
|n/a
|
|
 +
|-
 +
|2
 +
|J61100 + ladA
 +
|1625
 +
|<font color=limegreen>✓</font>
|
|
|-
|-
-
|2-6
+
|3
|J61100 + ladA
|J61100 + ladA
|1625
|1625
-
|2, 3, 4, 5
+
|<font color=limegreen>✓</font>
 +
|
|-
|-
-
|7-11
+
|4
 +
|J61100 + ladA
 +
|1625
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|5
 +
|J61100 + ladA
 +
|1625
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|6
 +
|J61100 + ladA
 +
|1625
 +
|<font color=red>✗</font>
 +
|
 +
|-
 +
|7
|J61101 + ADH
|J61101 + ADH
|1052
|1052
-
|7, 8, 10
+
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|8
 +
|J61101 + ADH
 +
|1052
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|9
 +
|J61101 + ADH
 +
|1052
 +
|<font color=red>✗</font>
 +
|
 +
|-
 +
|10
 +
|J61101 + ADH
 +
|1052
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|11
 +
|J61101 + ADH
 +
|1052
 +
|<font color=red>✗</font>
 +
|
|-
|-
-
|12-16
+
|12
|J61107 + ALDH
|J61107 + ALDH
|1796
|1796
 +
|<font color=red>✗</font>
 +
|
 +
|-
|13
|13
 +
|J61107 + ALDH
 +
|1796
 +
|<font color=limegreen>✓</font>
 +
|
 +
|-
 +
|14
 +
|J61107 + ALDH
 +
|1796
 +
|<font color=red>✗</font>
 +
|
 +
|-
 +
|15
 +
|J61107 + ALDH
 +
|1796
 +
|<font color=red>✗</font>
 +
|
 +
|-
 +
|16
 +
|J61107 + ALDH
 +
|1796
 +
|<font color=red>✗</font>
 +
|
|-
|-
|17
|17
|J61100 (uncut)
|J61100 (uncut)
-
|various
+
|
 +
|<font color=limegreen>✓</font>
|
|
|-
|-
|18
|18
|J61101 (uncut)
|J61101 (uncut)
-
|various
+
|
 +
|<font color=limegreen>✓</font>
|
|
|-
|-
|19
|19
|J61107 (uncut)
|J61107 (uncut)
-
|various
+
|
 +
|<font color=limegreen>✓</font>
|
|
|-
|-
|20
|20
-
|SmartLadder (5 μL)
+
|SmartLadder
 +
|n/a
 +
|n/a
|
|
-
|
 
-
|-
 
|}
|}
From the scPCR it can be concluded that from all the colonies PCRed (per BioBrick), at least one was positive! Next week the next (last) step in making the BioBricks for alkane degradation can be be done, so hopefully the week after that we can start characterizing.
From the scPCR it can be concluded that from all the colonies PCRed (per BioBrick), at least one was positive! Next week the next (last) step in making the BioBricks for alkane degradation can be be done, so hopefully the week after that we can start characterizing.

Revision as of 19:57, 2 August 2010

Alkane degradation

The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a colony PCR was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.

1% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker

Lane Description part 1

# Description Expected lenght (bp) Status Remarks
1 SmartLadder n/a n/a
2 J61100 + rubA3 473 ✓
3 J61100 + rubA3 473 ✓
4 J61100 + rubA3 473 ✗
5 J61100 + rubA3 473 ✓
6 J61100 + rubA3 473 ✗
7 J61100 + rubA4 482 ✓
8 J61100 + rubA4 482 ✓
9 J61100 + rubA4 482 ✓
10 J61100 + rubA4 482 ✓
11 J61100 + rubA4 482 ✓
12 J61100 + rubR 1505 ✓
13 J61100 + rubR 1505 ✓
14 J61100 + rubR 1505 ✓
15 J61100 + rubR 1505 ✓
16 J61100 + rubR 1505 ✓
17 SmartLadder n/a n/a
% agarose of colony PCR part 1. Gel runned at 100V for 1 hour. Of all samples 10 μL + 2 μL loadingbuffer was loaded. 5 μL was loaded of marker

Lane Description part 2

# Description Expected lenght (bp) Status Remarks
1 SmartLadder n/a n/a
2 J61100 + ladA 1625 ✓
3 J61100 + ladA 1625 ✓
4 J61100 + ladA 1625 ✓
5 J61100 + ladA 1625 ✓
6 J61100 + ladA 1625 ✗
7 J61101 + ADH 1052 ✓
8 J61101 + ADH 1052 ✓
9 J61101 + ADH 1052 ✗
10 J61101 + ADH 1052 ✓
11 J61101 + ADH 1052 ✗
12 J61107 + ALDH 1796 ✗
13 J61107 + ALDH 1796 ✓
14 J61107 + ALDH 1796 ✗
15 J61107 + ALDH 1796 ✗
16 J61107 + ALDH 1796 ✗
17 J61100 (uncut) ✓
18 J61101 (uncut) ✓
19 J61107 (uncut) ✓
20 SmartLadder n/a n/a

From the scPCR it can be concluded that from all the colonies PCRed (per BioBrick), at least one was positive! Next week the next (last) step in making the BioBricks for alkane degradation can be be done, so hopefully the week after that we can start characterizing.

Retrieved from "http://2010.igem.org/Team:TU_Delft/28_July_2010_content"