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 [https://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- | + | A [[Team:TU_Delft/protocols/transformation|transformation]] of [https://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== | ||
- | < | + | <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 [https://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| | + | [[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 | + | |SmartLadder |
- | | | + | |n/a |
- | | | + | |n/a |
+ | |n/a | ||
+ | | | ||
|- | |- | ||
|2 | |2 | ||
- | |J23100-B0030-I13401 | + | |transformant #1 of ligation mix: J23100-B0030-I13401 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|3 | |3 | ||
- | |J23100-B0030-I13401 | + | |transformant #2 of ligation mix: J23100-B0030-I13401 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|4 | |4 | ||
- | |J23100-B0030-I13401 | + | |transformant #3 of ligation mix: J23100-B0030-I13401 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|5 | |5 | ||
- | |J23100-B0030-I13401 | + | |transformant #4 of ligation mix: J23100-B0030-I13401 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|6 | |6 | ||
- | |J23100-B0030-I13401 | + | |transformant #5 of ligation mix: J23100-B0030-I13401 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|7 | |7 | ||
- | |I13401 ligation control | + | |transformant #1 of ligation mix: I13401 ligation control |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | |<font color=red>✖</font> | ||
| | | | ||
- | |||
|- | |- | ||
|8 | |8 | ||
- | |I13401 ligation control | + | |transformant #2 of ligation mix: I13401 ligation control |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|9 | |9 | ||
- | |I13401 ligation control | + | |transformant #3 of ligation mix: I13401 ligation control |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|10 | |10 | ||
- | |I13401 digestion control | + | |transformant #1 of ligation mix: I13401 digestion control |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|11 | |11 | ||
- | |I13401 digestion control | + | |transformant #2 of ligation mix: I13401 digestion control |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|12 | |12 | ||
- | | | + | |transformant #1 of K081005 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|13 | |13 | ||
- | | | + | |transformant #2 of K081005 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|14 | |14 | ||
- | | | + | |transformant #3 of K081005 |
+ | | | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|15 | |15 | ||
|BioRad EZ Load | |BioRad EZ Load | ||
- | | | + | |n/a |
- | | | + | |n/a |
+ | |n/a | ||
+ | | | ||
|- | |- | ||
|16 | |16 | ||
|PCR product of E0240 | |PCR product of E0240 | ||
+ | |1114 | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|17 | |17 | ||
|PCR product of E0240 | |PCR product of E0240 | ||
+ | |1114 | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|- | |- | ||
|18 | |18 | ||
|PCR product of E0240 | |PCR product of E0240 | ||
+ | |1114 | ||
+ | |G00101 + G00101 | ||
+ | ||<font color=limegreen>✔</font> | ||
| | | | ||
- | |||
|} | |} |
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)
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 | ✔ |