Team:TU Delft/22 July 2010 content

From 2010.igem.org

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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.
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==Salt tolerance==
 
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Time to get cracking, so bbc1 and B0015 were restricted using the scheme shown below.
 
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
 
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|'''#'''
 
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|'''Sample added'''
 
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|'''Volume added (µl)'''
 
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|1
 
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|bbc1
 
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|3
 
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|Ecro R1
 
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|1
 
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|Spe1
 
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|1
 
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|Buffer 2 (biolabs)
 
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|2
 
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|Water
 
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|11
 
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|BSA
 
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|2
 
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|2
 
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|B0015
 
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|3
 
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|-
 
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|Ecro R1
 
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|1
 
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|Xba1
 
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|1
 
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|-
 
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|Buffer 2 (biolabs)
 
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|2
 
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|-
 
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|Water
 
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|11
 
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|BSA
 
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|2
 
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|}
 

Revision as of 09:23, 10 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
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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