Team:TU Delft/22 July 2010 content

From 2010.igem.org

(Difference between revisions)
(→Alkane degradation)
(→Salt tolerance)
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Time to get cracking, so bbc1 and B0015 were restricted using the scheme shown below.
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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|-
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|1
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|bbc1
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|3
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|-
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|
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|Ecro R1
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|1
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|-
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|
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|Spe1
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|1
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|-
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|
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|Buffer 2 (biolabs)
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|2
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|-
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|
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|Water
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|11
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|-
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|
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|BSA
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|2
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|-
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|2
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|B0015
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|3
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|-
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|
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|Ecro R1
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|1
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|-
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|
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|Xba1
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|1
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|-
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|
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|Buffer 2 (biolabs)
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|2
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|-
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|
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|Water
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|11
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|-
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|
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|BSA
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|2
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|}

Revision as of 09:15, 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.


Salt tolerance

Time to get cracking, so bbc1 and B0015 were restricted using the scheme shown below.

# Sample added Volume added (µl)
1 bbc1 3
Ecro R1 1
Spe1 1
Buffer 2 (biolabs) 2
Water 11
BSA 2
2 B0015 3
Ecro R1 1
Xba1 1
Buffer 2 (biolabs) 2
Water 11
BSA 2
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