Team:TU Delft/20 July 2010 content

From 2010.igem.org

(Difference between revisions)
(Characterization of Anderson RBS sequences)
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 +
=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 [[Team:TU_Delft/protocols/making_competent_cells|competent cells]]
 +
==Alkane degradation==
==Alkane degradation==
'''BioBrick production continued'''
'''BioBrick production continued'''
-
A [[Team:TU_Delft/protocols/transformation|transformation]] of [http://2010.igem.org/Team:TU_Delft#/blog?blog=19_July_2010 yesterday's] ligation products was done in TOP10 competent cells, and grown on LB-TET plates overnight. Hopefully we will have some colonies tomorrow.
+
A [[Team:TU_Delft/protocols/transformation|transformation]] of [http://2010.igem.org/Team:TU_Delft#/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.
==Characterization of Anderson RBS sequences==
==Characterization of Anderson RBS sequences==
-
<h5>Assembly of reference construct</h5>
+
<h4>Assembly of reference construct</h4>
-
Single colony PCRs were performed on the transformants obtained yesterday. These were loaded onto 1% agarose gel (see gel below)
+
Single [[Team:TU_Delft/protocols/Colony_PCR|colony PCRs]] were performed on the transformants obtained [http://2010.igem.org/Team:TU_Delft#/blog?blog=19_July_2010 yesterday]. These were loaded onto [[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] (see gel below)
-
[[Image:TU_Delft_ScPCR-digPCR-20Jul10.png|500px|thumb|left|scPCR]]
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[[Image:TU_Delft_ScPCR-digPCR-20Jul10.png|500px|thumb|left|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:
Lane description:
Line 16: Line 24:
|'''Description'''
|'''Description'''
|'''Expected Length (bp)'''
|'''Expected Length (bp)'''
 +
|'''Primers'''
 +
|'''Status'''
|'''Remarks'''
|'''Remarks'''
|-
|-
|1
|1
-
|SmartLadder (5 μL)
+
|SmartLadder
-
|Varying
+
|n/a
-
|Visible
+
|n/a
 +
|n/a
 +
|
|-
|-
|2
|2
-
|J23100-B0030-I13401 colony 1
+
|transformant #1 of ligation mix: J23100-B0030-I13401
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|3
|3
-
|J23100-B0030-I13401 colony 2
+
|transformant #2 of ligation mix: J23100-B0030-I13401
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|4
|4
-
|J23100-B0030-I13401 colony 3
+
|transformant #3 of ligation mix: J23100-B0030-I13401
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|5
|5
-
|J23100-B0030-I13401 colony 4
+
|transformant #4 of ligation mix: J23100-B0030-I13401
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|6
|6
-
|J23100-B0030-I13401 colony 5
+
|transformant #5 of ligation mix: J23100-B0030-I13401
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|7
|7
-
|I13401 ligation control colony 1
+
|transformant #1 of ligation mix: I13401 ligation control  
 +
|
 +
|G00101 + G00101
 +
|<font color=red>&#10006;</font>
|
|
-
|Not visible
 
|-
|-
|8
|8
-
|I13401 ligation control colony 2
+
|transformant #2 of ligation mix: I13401 ligation control  
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|9
|9
-
|I13401 ligation control colony 3
+
|transformant #3 of ligation mix: I13401 ligation control  
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Not visible
 
|-
|-
|10
|10
-
|I13401 digestion control colony 1
+
|transformant #1 of ligation mix: I13401 digestion control
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|11
|11
-
|I13401 digestion control colony 2
+
|transformant #2 of ligation mix: I13401 digestion control
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|12
|12
-
|K081005 colony 1
+
|transformant #1 of K081005
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|13
|13
-
|K081005 colony 2
+
|transformant #2 of K081005
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|14
|14
-
|K081005 colony 3
+
|transformant #3 of K081005
 +
|
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|15
|15
|BioRad EZ Load
|BioRad EZ Load
-
|Varying
+
|n/a
-
|Visible
+
|n/a
 +
|n/a
 +
|
|-
|-
|16
|16
|PCR product of E0240
|PCR product of E0240
 +
|1114
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|17
|17
|PCR product of E0240
|PCR product of E0240
 +
|1114
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|-
|-
|18
|18
|PCR product of E0240
|PCR product of E0240
 +
|1114
 +
|G00101 + G00101
 +
||<font color=limegreen>&#10004;</font>
|
|
-
|Visible
 
|}
|}

Revision as of 18:59, 1 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.

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