Team:Paris Saclay/Notebook/July/21

From 2014.igem.org

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(3 - Liquid Bacterial Culture)
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{{Team:Paris_Saclay/notebook_header}}
=Monday 21st July=
=Monday 21st July=
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==Lab Work==
==Lab Work==
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===The chassis coli Odor free===
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===1 - Results: Transformation of supercompetent cells with CaCl2===
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====Results: Transformation of CaCl<sub>2</sub> supercompetent cells====
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''by Romain''
''by Romain''
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from Bacterial Cultures transformed the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July]
from Bacterial Cultures transformed the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July]
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Results: Nothing has grown.
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'''Results:''' Nothing has grown.
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===2 - Oligo's design===
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====Results: Transformation of DY330 via pJBEI6409====
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''by Sean''
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''by Romain''
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Transformation performed on the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July]
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===3 - Liquid Bacterial Culture===
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{| class="wikitable"
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|+Number of colonies per dish (source of bacteria: electroporation cuvettes prepared on the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July])
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|-
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! Volume of plasmid used
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! 50μl from cuvette
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! 100μl
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! remainder (850µl)
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|-
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! 2μl of plasmid
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| 0
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| 4
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| 30
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|-
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! 4μl
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| 3
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| 6
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| 45
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|-
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! Control
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| 0
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| 0
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| 0
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|}
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'''Conclusion:''' The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.
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===Lemon scent===
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====Liquid Bacterial Culture====
''by Marie, Romain & Sean''
''by Marie, Romain & Sean''
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*The new strains received
*The new strains received
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[https://2014.igem.org/Team:Paris_Saclay/Notebook Back to the Calendar]
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====Electrophoresis====
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''by Fabio (process A) and Mathieu (process B and C)''
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 +
[[File:LU000069.jpg|right]]
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[[File:LU000071.jpg|right]]
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[[File:LU000070.jpg|right]]
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 +
We used 2µl of DNA of the following Biobricks in a 1% Agarose Gel.
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''The PCRs came from Mathias' manipulation made the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July].''
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'''''Process A'''''
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 BBa_J23119 Cl1]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 BBa_J23119 Cl2]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23106 Cl1]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23100 Cl2]
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#PCR 1
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#PCR 2
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#PCR 3
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#PCR 4
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#PCR 5
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#PCR 6
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#PCR 7
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#PCR 8
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#PCR 9
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#PCR 10
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'''''Process B'''''
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23100 Cl1]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23100 Cl2]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23106 Cl1]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23106 Cl2]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23114 Cl1]
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#[https://2014.igem.org/Team:Paris_Saclay/Notebook/July/16 BBa_J23114 Cl2]
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'''''Process C'''''
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''Pooling and purifying PCR 9 and 10 from process A.''
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#PCR purified products. IPS70 / IPS71 of pOsV230 (Apramycin)
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#BT 340 (plasmid's flipase)
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'''Results:'''
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*'''A''': From 1 to 4: Success, DNAs have the expected size.
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*'''A''': From 5 to 12: Failure, No PCR products.
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*'''A''': Numbers 13 and 14: Success, PCR products have the expected size.
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*'''B''': All 6 extractions were successful.
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*'''C''': Number 1: successful concentration of pOsV230's PCR product.
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*'''C''': Number 2 had no migration.
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Gel_Electrophoresis Protocol]
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==Photo of the Day==
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[[File:Paris Saclay 21_july.jpg|400px|center]]
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'''People there''':
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* Instructors and advisors: Solenne and Sylvie.
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* Students: Arnaud, Fabio, Juliette, Mathieu, Marie, Pierre, Romain, Sean and Terry.
 +
 
 +
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 15:24, 14 October 2014

Contents

Monday 21st July

Lab Work

The chassis coli Odor free

Results: Transformation of CaCl2 supercompetent cells

by Romain

Strains transformed: MG1655, MG1655Z1. from Bacterial Cultures transformed the 18th July

Results: Nothing has grown.

Results: Transformation of DY330 via pJBEI6409

by Sean

Transformation performed on the 18th July

Number of colonies per dish (source of bacteria: electroporation cuvettes prepared on the 18th July)
Volume of plasmid used 50μl from cuvette 100μl remainder (850µl)
2μl of plasmid 0 4 30
4μl 3 6 45
Control 0 0 0

Conclusion: The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.

Lemon scent

Liquid Bacterial Culture

by Marie, Romain & Sean

  • DY330 pJBEI6409 with 10µl Cm in 10ml LB (x2)
  • BT340 Cm and Amp
  • The new strains received

Electrophoresis

by Fabio (process A) and Mathieu (process B and C)

LU000069.jpg
LU000071.jpg
LU000070.jpg

We used 2µl of DNA of the following Biobricks in a 1% Agarose Gel. The PCRs came from Mathias' manipulation made the 18th July.

Process A

  1. BBa_J23119 Cl1
  2. BBa_J23119 Cl2
  3. BBa_J23106 Cl1
  4. BBa_J23100 Cl2
  5. PCR 1
  6. PCR 2
  7. PCR 3
  8. PCR 4
  9. PCR 5
  10. PCR 6
  11. PCR 7
  12. PCR 8
  13. PCR 9
  14. PCR 10

Process B

  1. BBa_J23100 Cl1
  2. BBa_J23100 Cl2
  3. BBa_J23106 Cl1
  4. BBa_J23106 Cl2
  5. BBa_J23114 Cl1
  6. BBa_J23114 Cl2

Process C

Pooling and purifying PCR 9 and 10 from process A.

  1. PCR purified products. IPS70 / IPS71 of pOsV230 (Apramycin)
  2. BT 340 (plasmid's flipase)

Results:

  • A: From 1 to 4: Success, DNAs have the expected size.
  • A: From 5 to 12: Failure, No PCR products.
  • A: Numbers 13 and 14: Success, PCR products have the expected size.
  • B: All 6 extractions were successful.
  • C: Number 1: successful concentration of pOsV230's PCR product.
  • C: Number 2 had no migration.

Protocol

Photo of the Day

Paris Saclay 21 july.jpg

People there:

  • Instructors and advisors: Solenne and Sylvie.
  • Students: Arnaud, Fabio, Juliette, Mathieu, Marie, Pierre, Romain, Sean and Terry.

Back to the calendar