Team:TU Delft/20 July 2010 content

From 2010.igem.org

(Difference between revisions)
(Assembly of reference construct)
(Alkane degradation)
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A [[Team:TU_Delft/protocols/transformation|transformation]] of [http://2010.igem.org/Team:TU_Delft#page=pages/blog&blog=19_July_2010 yesterday's] ligation products was done in TOP10 competent cells, and grown on LB-Tetracycline plates overnight. Hopefully we will have some colonies tomorrow.
A [[Team:TU_Delft/protocols/transformation|transformation]] of [http://2010.igem.org/Team:TU_Delft#page=pages/blog&blog=19_July_2010 yesterday's] ligation products was done in TOP10 competent cells, and grown on LB-Tetracycline plates overnight. Hopefully we will have some colonies tomorrow.
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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. These were then ligated to one another in the third scheme.
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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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|-
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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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|-
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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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|Ecro R1
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|1
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|-
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|Xba1
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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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|-
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|
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|BSA
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|2
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|-
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|3
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|B0015 (Cut)
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|1
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|-
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|Ecro R1
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|1
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|-
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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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|Water
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|11
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==Characterization of Anderson RBS sequences==
==Characterization of Anderson RBS sequences==

Revision as of 09:25, 10 August 2010

Contents

Juggling an Diabolo tricks!

From all the pipetting that we do in the lab, our fingers and wrists are a bit tense. After some hard thinking, we found a perfect way to relax them…..by juggling and performing diabolo tricks! So now, in between the incubation times of our experiments the juggle balls are flying around in our iGEM room.

Lab work

Competent cells

Eva made another batch of competent cells

Alkane degradation

BioBrick production continued

A transformation of yesterday's ligation products was done in TOP10 competent cells, and grown on LB-Tetracycline plates overnight. Hopefully we will have some colonies tomorrow.


Salt tolerance

Time to get cracking, so bbc1 and B0015 were restricted using the scheme shown below. These were then ligated to one another in the third scheme.

# 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
3 B0015 (Cut) 1
Ecro R1 1
Xba1 1
Buffer 2 (biolabs) 2
Water 11
BSA 2

Characterization of Anderson RBS sequences

Assembly of reference construct

Single colony PCRs were performed on the transformants obtained yesterday. These were loaded onto 1% agarose gel (see gel below)

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

Lane description:

# Description Expected Length (bp) Primers Status Remarks
1 SmartLadder n/a n/a n/a
2 transformant #1 of ligation mix: J23100-B0030-I13401 G00101 + G00101
3 transformant #2 of ligation mix: J23100-B0030-I13401 G00101 + G00101
4 transformant #3 of ligation mix: J23100-B0030-I13401 G00101 + G00101
5 transformant #4 of ligation mix: J23100-B0030-I13401 G00101 + G00101
6 transformant #5 of ligation mix: J23100-B0030-I13401 G00101 + G00101
7 transformant #1 of ligation mix: I13401 ligation control G00101 + G00101
8 transformant #2 of ligation mix: I13401 ligation control G00101 + G00101
9 transformant #3 of ligation mix: I13401 ligation control G00101 + G00101
10 transformant #1 of ligation mix: I13401 digestion control G00101 + G00101
11 transformant #2 of ligation mix: I13401 digestion control G00101 + G00101
12 transformant #1 of K081005 G00101 + G00101
13 transformant #2 of K081005 G00101 + G00101
14 transformant #3 of K081005 G00101 + G00101
15 BioRad EZ Load n/a n/a n/a
16 PCR product of E0240 1114 G00101 + G00101
17 PCR product of E0240 1114 G00101 + G00101
18 PCR product of E0240 1114 G00101 + G00101

Beach

After a hard day working in the lab, we went to the beach in Scheveningen for a nice walk and to watch the sunset.