Team:TU Delft/22 July 2010 content

From 2010.igem.org

(Difference between revisions)
(→Alkane degradation)
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=Lab work=
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==Alkane degradation==
==Alkane degradation==
There were some colonies on [https://2010.igem.org/Team:TU_Delft#/blog?blog=19_July_2010 Tuesday's] plates! We had left the plates @ 37°C yesterday after having seen that there were no colonies. When checking this morning on all plates (except the negative control) there were a few colonies! (2-50 colonies). Chances are it's not what we're looking for, but maybe they are good transformants... to check we will do a [[Team:TU_Delft/protocols/colony PCR|colony PCR]].
There were some colonies on [https://2010.igem.org/Team:TU_Delft#/blog?blog=19_July_2010 Tuesday's] plates! We had left the plates @ 37°C yesterday after having seen that there were no colonies. When checking this morning on all plates (except the negative control) there were a few colonies! (2-50 colonies). Chances are it's not what we're looking for, but maybe they are good transformants... to check we will do a [[Team:TU_Delft/protocols/colony PCR|colony PCR]].
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[[Image:TUDelft_scPCR_22-07.png|550px|thumb|left|1% Agarose gel of SC PCR samples]]
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[[Image:TUDelft_scPCR_22-07.png|550px|thumb|left|1% Agarose gel of colony PCR. Gel runned at 100V for 1 hour. Of all samples 5 μL + 1 μL loadingbuffer was loaded. 5 μL was loaded of marker.]]
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
|'''#'''
|'''#'''
|'''Description'''
|'''Description'''
|'''Expected length (bp)'''
|'''Expected length (bp)'''
 +
|'''Primers'''
 +
|'''Status'''
 +
|'''Remarks'''
|-
|-
|1
|1
-
|[https://2010.igem.org/Image:TU_Delft_SmartLadder.jpg SmartLadder] (5 μL)
+
|[https://2010.igem.org/Image:TU_Delft_SmartLadder.jpg SmartLadder]
|n/a
|n/a
 +
|n/a
 +
|n/a
 +
|
|-
|-
-
|2-4
+
|2
-
|007T
+
|Transformant #1 of ligation mix 007T
|1616
|1616
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
-
|5-6
+
|3
-
|008T
+
|Transformant #2 of ligation mix 007T
-
|551
+
|1616
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|4
 +
|Transformant #3 of ligation mix 007T
 +
|1616
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|5
 +
|Transformant #1 of ligation mix 008T
 +
|551
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|6
 +
|Transformant #2 of ligation mix 008T
 +
|551  
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|7
 +
|Transformant #1 of ligation mix 009T
 +
|551
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
-
|7-9
+
|8
-
|009T
+
|Transformant #1 of ligation mix 009T
|560
|560
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
-
|10-12
+
|9
-
|010T
+
|Transformant #1 of ligation mix 009T
 +
|560
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|10
 +
|Transformant #1 of ligation mix 010T
 +
|1657
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|11
 +
|Transformant #1 of ligation mix 010T
 +
|1657
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|12
 +
|Transformant #1 of ligation mix 010T
|1657
|1657
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
-
|13-15
+
|13
-
|017T
+
|Transformant #1 of ligation mix 017T
|1130
|1130
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|14
 +
|Transformant #1 of ligation mix 017T
 +
|1130
 +
|G00100 + G00101
 +
|
 +
|
 +
|-
 +
|15
 +
|Transformant #1 of ligation mix 017T
 +
|1130
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
|16
|16
-
|018T
+
|Transformant #1 of ligation mix 018T
|1874
|1874
 +
|G00100 + G00101
 +
|
 +
|
|-
|-
|17
|17
-
|Red colony
+
|Transformant #1 of Red colony
-
|n/a
+
|1360
 +
|G00100 + G00101
 +
|
 +
|
|}
|}
A number of colonies look promising! To check if they really are the BioBricks we want, tomorrow we will do a plasmid isolation with the cultures of lane 2, 4, 6, 7, 9 and 14. We will cut the isolated plasmids with various restriction enzymes and analyze the digestion products on gel.
A number of colonies look promising! To check if they really are the BioBricks we want, tomorrow we will do a plasmid isolation with the cultures of lane 2, 4, 6, 7, 9 and 14. We will cut the isolated plasmids with various restriction enzymes and analyze the digestion products on gel.

Revision as of 19:25, 1 August 2010

Lab work

Alkane degradation

There were some colonies on Tuesday's plates! We had left the plates @ 37°C yesterday after having seen that there were no colonies. When checking this morning on all plates (except the negative control) there were a few colonies! (2-50 colonies). Chances are it's not what we're looking for, but maybe they are good transformants... to check we will do a colony PCR.

1% Agarose gel of colony PCR. Gel runned at 100V for 1 hour. Of all samples 5 μL + 1 μL loadingbuffer was loaded. 5 μL was loaded of marker.
# Description Expected length (bp) Primers Status Remarks
1 SmartLadder n/a n/a n/a
2 Transformant #1 of ligation mix 007T 1616 G00100 + G00101
3 Transformant #2 of ligation mix 007T 1616 G00100 + G00101
4 Transformant #3 of ligation mix 007T 1616 G00100 + G00101
5 Transformant #1 of ligation mix 008T 551 G00100 + G00101
6 Transformant #2 of ligation mix 008T 551 G00100 + G00101
7 Transformant #1 of ligation mix 009T 551 G00100 + G00101
8 Transformant #1 of ligation mix 009T 560 G00100 + G00101
9 Transformant #1 of ligation mix 009T 560 G00100 + G00101
10 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
11 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
12 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
13 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
14 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
15 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
16 Transformant #1 of ligation mix 018T 1874 G00100 + G00101
17 Transformant #1 of Red colony 1360 G00100 + G00101

A number of colonies look promising! To check if they really are the BioBricks we want, tomorrow we will do a plasmid isolation with the cultures of lane 2, 4, 6, 7, 9 and 14. We will cut the isolated plasmids with various restriction enzymes and analyze the digestion products on gel.

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