Difference between revisions of "Team:TU Darmstadt/Notebook"

(reporterkram rein)
(reporter verbessert)
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976002:</b><br>
 
<b>BBa&#95;K1976002:</b><br>
−
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones. </p>
+
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones. <br>
  
−
</td></tr>
 
  
−
<td><p><h6>Metabolic burden</h6>
+
<h6>Metabolic burden</h6>
 
<b>BBa&#95;K1976001:</b><br>
 
<b>BBa&#95;K1976001:</b><br>
 
The BioBrick BBa&#95;I11020 was synthezised with a LVA&#8209;tag and with B0034 as a ribosome binding site by IDT.</p>
 
The BioBrick BBa&#95;I11020 was synthezised with a LVA&#8209;tag and with B0034 as a ribosome binding site by IDT.</p>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976002:</b><br>
 
<b>BBa&#95;K1976002:</b><br>
−
Isolated Plasmid DNA Sequence was verified by Sanger Sequencing. </p>
+
Isolated Plasmid DNA Sequence was verified by Sanger Sequencing. <br>
−
</td></tr>
+
<h6>Metabolic burden</h6>
−
<td><p><h6>Metabolic burden</h6>
+
 
<b>BBa&#95;K1976001:</b><br>
 
<b>BBa&#95;K1976001:</b><br>
 
The LVA&#8209;tag was deleted by PCR with the primer Del&#95;LVA&#95;fw and Del&#95;LVA&#95;rev.</p>
 
The LVA&#8209;tag was deleted by PCR with the primer Del&#95;LVA&#95;fw and Del&#95;LVA&#95;rev.</p>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976003:</b><br>
 
<b>BBa&#95;K1976003:</b><br>
−
The mVenus-LVA-pSB1C3 plasmid was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a T7-RBS-pSB1A3 plasmid which was cut with <i>SpeI</i> and <i>PstI.</i> </p>
+
The mVenus-LVA-pSB1C3 plasmid was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a T7-RBS-pSB1A3 plasmid which was cut with <i>SpeI</i> and <i>PstI.</i> <br>
−
</td></tr>
+
  
−
<td><p><h6>Metabolic burden</h6>
+
<h6>Metabolic burden</h6>
  
 
<b>BBa&#95;K1976000:</b><br>
 
<b>BBa&#95;K1976000:</b><br>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976003:</b><br>
 
<b>BBa&#95;K1976003:</b><br>
−
The ligation product was transformed into <i>E.coli</i> Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate. </p>
+
The ligation product was transformed into <i>E.coli</i> Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate. <br>
−
</td></tr>
+
  
  
−
<td><p><h6>Metabolic burden</h6>
+
<h6>Metabolic burden</h6>
  
 
<b>BBa&#95;K1976000:</b>
 
<b>BBa&#95;K1976000:</b>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976003:</b><br>
 
<b>BBa&#95;K1976003:</b><br>
−
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones . </p>
+
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones . <br>
  
−
</td></tr>
+
 
−
<td><p><h6>Metabolic burden</h6>
+
<h6>Metabolic burden</h6>
  
 
<b>BBa&#95;K1976000:</b><br>
 
<b>BBa&#95;K1976000:</b><br>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976003:</b><br>
 
<b>BBa&#95;K1976003:</b><br>
−
The isolated Plasmid DNA was verified by Sanger sequencing.</p>
+
The isolated Plasmid DNA was verified by Sanger sequencing.<br>
−
</td></tr>
+
<h6>Metabolic burden</h6>
−
<td><p><h6>Metabolic burden</h6>
+
  
 
<b>BBa&#95;K1976000:</b><br>
 
<b>BBa&#95;K1976000:</b><br>
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<td><p><h6>Reporter System</h6>
 
<td><p><h6>Reporter System</h6>
 
<b>BBa&#95;K1976003:</b><br>
 
<b>BBa&#95;K1976003:</b><br>
−
Subsequently the psB1A3 vector containing brick was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a pSB1C3 vector using T4&#8209;Ligase.</p>
+
Subsequently the psB1A3 vector containing brick was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a pSB1C3 vector using T4&#8209;Ligase.<br>
−
</td></tr>
+
<h6>Metabolic burden</h6>
−
<td><p><h6>Metabolic burden</h6>
+
  
 
<b>BBa&#95;K1976000:</b><br>
 
<b>BBa&#95;K1976000:</b><br>
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<div class="labWeek"><h5>Lab Week 27: <div class="week_date"> October 17 - October 23</div></h5></div></td>
 
<div class="labWeek"><h5>Lab Week 27: <div class="week_date"> October 17 - October 23</div></h5></div></td>
 
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td>
 
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td>
−
<td><p><h6>Metabolic burden</h6>
+
<p><h6>Metabolic burden</h6>
 
Plasmid curing could not be performed due to lack of time.
 
Plasmid curing could not be performed due to lack of time.
 
</p>
 
</p>

Revision as of 17:28, 19 October 2016

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iGEM TU Darmstadt 2016
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NOTEBOOK

Labjournal 2016

For half a year we were working in the wet lab    , developed a pipetting robot   and simulated the molecular dynamics of our proteins    . As well we would like to highlight certain social events   which helped us to further improve our team spirit.

Lab Week 1:
April 18 - April 24

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 2:
April 25 - Mai 1

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 3:
May 2 - May 8

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 4:
May 9 - May 15

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 5:
May 16 - May 22

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 6:
May 23 - May 29

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 7:
May 30 - June 5

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 8:
June 6 - June 12

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 9:
June 13 - June 19

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 10:
June 20 - June 26

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 11:
June 27 - July 3

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 12:
July 4 - July 10

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 13:
July 11 - July 17

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 14:
July 18 - July 24

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 15:
July 25 - July 31

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 16:
August 1 - August 7

Hier könnte Ihr labjournal stehen!

robotics
modeling
social
Lab Week 17:
August 8 - August 14

Reporter System
BBa_K1976002:
The mVenus-LVA-gene was synthesized by idt and amplified through PCR using the SP-amplify-fw-Rep1 (fw) and SP-amplify-rev-Rep2 (REV) oligonucleotides.

robotics
modeling
social
Lab Week 18:
August 15 - August 21

Reporter System
BBa_K1976002:
The PCR product was verified by agarose gel electrophoresis and subsequently purified. The mVenus amplicon was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4-ligase.

robotics
modeling
social
Lab Week 19:
August 22 - August 28

Reporter System
BBa_K1976002:
The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a CMP agar plate.

robotics
modeling
social
Lab Week 20:
August 29 - September 4

Reporter System
BBa_K1976002:
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones.
Metabolic burden
BBa_K1976001:
The BioBrick BBa_I11020 was synthezised with a LVA‑tag and with B0034 as a ribosome binding site by IDT.

robotics
modeling
social
Lab Week 21:
September 5 - September 11

Reporter System
BBa_K1976002:
Isolated Plasmid DNA Sequence was verified by Sanger Sequencing.
Metabolic burden
BBa_K1976001:
The LVA‑tag was deleted by PCR with the primer Del_LVA_fw and Del_LVA_rev.

robotics
modeling
social
Lab Week 22:
September 12 - September 18

Reporter System
BBa_K1976003:
The mVenus-LVA-pSB1C3 plasmid was cut with EcoRI and PstI and ligated into a T7-RBS-pSB1A3 plasmid which was cut with SpeI and PstI.
Metabolic burden
BBa_K1976000:
The BioBrick BBa_E0240 was cut with the restriction enzymes XbaI and PstI.
BBa_J61002 was cut with SpeI and PstI.
The two restriction products were ligated by standard T4‑Ligation protocol.
BBa_K1976001:
B0034‑BBa_I11020 was cut with XbaI and PstI.
BBa_J23101 was cut with SpeI and PstI.
Both fragments were ligated using the T4‑Ligase by standard protocol.

robotics
modeling
social
Lab Week 23:
September 19 - September 25

Reporter System
BBa_K1976003:
The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate.
Metabolic burden
BBa_K1976000:
After plasmid preparation, the plasmid was cut with XbaI and PstI.
We had the attp‑site BBa_11023 flanked by the two additional bidirectional transcription terminators BBa_1001 synthesized by IDT. The attp‑site of BBa_11023 was not the original λ‑attp‑site. Instead the sequence corresponded to the attP2‑site used in the Gateway ® cloning system.
The construct was cut with EcoRI and SpeI. The pSB1C3 backbone was cut with EcoRI and PstI. Together with the previously cut BBa_J61002‑BBa_E0240 construct the synthesized BBa_1001‑BBa_11023‑BBa_1001 construct was ligated in the cut pSB1C3.
BBa_K1976001:
The ligation product was transformed into E. coli Top 10 via heatshock.

robotics
modeling
social
Lab Week 24:
September 26 - October 2

Reporter System
BBa_K1976003:
Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones .
Metabolic burden
BBa_K1976000:
The attP2‑site was mutated to λ‑attp via Quick‑Change‑PCR, using the following primers: QC_attp_fw and QC_attp_rev (for more details see our primerlist).
BBa_K1976001:
To check whether BBa_I11020 was expressed properly a SDS‑Page was made.

robotics
modeling
social
Lab Week 25:
October 3 - October 9

Reporter System
BBa_K1976003:
The isolated Plasmid DNA was verified by Sanger sequencing.
Metabolic burden
BBa_K1976000:
The LVA‑tag was added by PCR with the overhang primer LVA_GFP_fw and LVA_GFP_rev.

robotics
modeling
social
Lab Week 26:
October 10 - October 16

Reporter System
BBa_K1976003:
Subsequently the psB1A3 vector containing brick was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4‑Ligase.
Metabolic burden
BBa_K1976000:
For plasmid curing the final plasmid construct and the low copy backbone pSB4A5 were cut with EcoRI and PstI and then ligated.
BBa_K1976001:
For plasmid curing the final plasmid construct and the low copy vector pSB4K5 were cut with EcoRI and PstI and then ligated.
BBa_K1976000 and BBa_K1976001 were cotransformed into E. coli Top 10.
To check whether the genomic integration was successful a cPCR with attb_fw and VR primers was made.

robotics
modeling
social

Metabolic burden
Plasmid curing could not be performed due to lack of time.

Lab Week 27:
October 17 - October 23
robotics
modeling
social