Difference between revisions of "Team:Paris Saclay/Notebook/July/27"

(Created page with "=Friday 27th July= ==Lab Work== ===New culture=== ''by Seong Koo'' Biobricks: * BBa_K1707005 * BBa_K1707006 * BBa_K1707007 ===Ligation=== ''by Pauline and Coralie'' ====K17070...")
 
(→‎Digestion + Purification on agarose gel)
 
(6 intermediate revisions by 3 users not shown)
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=Friday 27th July=
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{{Team:Paris_Saclay/notebook_header}}
 +
=Monday 27th July=
 
==Lab Work==
 
==Lab Work==
 
===New culture===
 
===New culture===
Line 12: Line 13:
 
''by Pauline and Coralie''
 
''by Pauline and Coralie''
  
−
====K1707008====
+
* BBa_K1707008
−
* 2µL Tp 10x
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** 2µL Tp 10x
−
* 1µL Ligase
+
** 1µL Ligase
−
* 11µL BBa_K1707003
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** 11µL BBa_K1707003
−
* 6µL BBa_J23101
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** 6µL BBa_J23101
  
−
====K1707010====
+
* BBa_K1707010
−
* 1µL Tp 10x
+
** 1µL Tp 10x
−
* 1µL Ligase
+
** 1µL Ligase
−
* 12µL BBa_K1399005
+
** 12µL BBa_K1399005
−
* 10µL BBa_B0015
+
** 10µL BBa_B0015
−
* 1µL H20
+
** 1µL H20
−
====K1707009====
+
−
* 2µL Tp 10x
+
−
* 2µL Ligase
+
−
* 10µL BBa_S03518
+
−
* 6µL BBa_R0051
+
  
−
====K1707004====
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* BBa_K1707009
−
* 3µL Tp 10x
+
** 2µL Tp 10x
−
* 2µL Ligase
+
** 2µL Ligase
−
* 8µL BBa_K1707003
+
** 10µL BBa_S03518
−
* 7µL BBa_R0051
+
** 6µL BBa_R0051
−
Agarose gel 1%, 110V
+
  
−
We cut bands with a scalpel
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* BBa_K1707004
 +
** 3µL Tp 10x
 +
** 2µL Ligase
 +
** 8µL BBa_K1707003
 +
** 7µL BBa_R0051
  
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===Purification===
+
 
−
''by Audrey''
+
===Transformation===
 +
''by Pauline and Coralie''
  
 
Biobricks:
 
Biobricks:
−
* BBa_K1707003
+
* BBa_K1707008
−
* BBa_K1399005
+
* BBa_K1707010
−
* BBa_B0015
+
* BBa_K1707009
−
* BBa_J23101
+
* BBa_K1707004
  
−
With PCR Clean-up/Gel extraction kit from Macherey-Nigel
 
  
−
===Quantification===
+
 
 +
===Digestion + Purification + Quantification===
 +
'' by Audrey''
  
 
Biobricks:
 
Biobricks:
−
* BBa_K1707003
+
* BBa_R0051
−
* BBa_K1399005
+
* BBa_K1707000
−
* BBa_B0015
+
−
* BBa_J23101
+
  
−
Agarose gel 1%
+
Digestion: as usual, with SpeI + PstI
−
Migration 110V
+
  
−
===Transformation===
+
Migration: Agarose gel 1%, migration 100V
−
''by Seong-Koo''
+
 
 +
===Plasmid extraction===
 +
'' by Pauline''
  
 
Biobricks:
 
Biobricks:
−
* BBa_K1707005
+
* BBa_K1707003 #4 et #5
−
* BBa_K1707006
+
 
−
* BBa_K1707007
+
With Plasmid DNA purification kit from Macherey-Nagel
 +
 
 +
===Digestion + Purification on agarose gel===
 +
'' by Pauline''
 +
 
 +
Biobricks:
 +
* BBa_K1707003 #4 and #5
 +
 
 +
Mix:
 +
* 10µL plasmid BBa_K1707003
 +
* 1µL XbaI
 +
* 1µL PstI
 +
* 2µL Buffer FastDigest
 +
* 6µL H2O
 +
 
 +
Purification on agarose gel 1%, migration 100V
 +
[[File:ParisSaclay 27.07.15-quantification+vérif.jpg|200px|center]]
 +
<html><p><i>Wells 2-3: Quantification, Wells 9-10: Verification of purification; from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707000#2, 3. BBa_50051, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. BBa_K1707003#1, 10. BBa_K1707003#2, 11. Empty 12. Empty</i></p></html>
 +
Cut with a scalpel
 +
Purification with the PCR Clean up kit from Macherey-Nagel
 +
 
 +
===New Culture===
 +
''by Johan''
 +
 
 +
Strains:
 +
* 1696
 +
* 1693
 +
* 1320
  
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===Soil experiment===
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in 150mL LB without any antibiotic
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'' by Seong Koo''
+
  
−
Plates observation and count of CFU:
 
−
* Strain 1696:
 
−
** Dilution 1: 424 CFU
 
−
** Dilution 0.1: 75 CFU
 
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** Dilution 0.01: 1 CFU
 
  
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* Strain 1693:
 
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** Dilution 1: 524 CFU
 
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** Dilution 0.1: 35 CFU
 
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** Dilution 0.01: 0 CFU
 
  
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* Strain 1320:
 
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** Dilution 1: 111 CFU
 
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** Dilution 0.1: 13 CFU
 
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** Dilution 0.01: 0 CFU
 
  
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We take 1g of contaminated soil and treat it like yesterday to spread it on specific plates
 
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Incubation ON, 37°C
 
  
  

Latest revision as of 21:20, 18 September 2015

Monday 27th July

Lab Work

New culture

by Seong Koo

Biobricks:

  • BBa_K1707005
  • BBa_K1707006
  • BBa_K1707007

Ligation

by Pauline and Coralie

  • BBa_K1707008
    • 2µL Tp 10x
    • 1µL Ligase
    • 11µL BBa_K1707003
    • 6µL BBa_J23101
  • BBa_K1707010
    • 1µL Tp 10x
    • 1µL Ligase
    • 12µL BBa_K1399005
    • 10µL BBa_B0015
    • 1µL H20
  • BBa_K1707009
    • 2µL Tp 10x
    • 2µL Ligase
    • 10µL BBa_S03518
    • 6µL BBa_R0051
  • BBa_K1707004
    • 3µL Tp 10x
    • 2µL Ligase
    • 8µL BBa_K1707003
    • 7µL BBa_R0051


Transformation

by Pauline and Coralie

Biobricks:

  • BBa_K1707008
  • BBa_K1707010
  • BBa_K1707009
  • BBa_K1707004


Digestion + Purification + Quantification

by Audrey

Biobricks:

  • BBa_R0051
  • BBa_K1707000

Digestion: as usual, with SpeI + PstI

Migration: Agarose gel 1%, migration 100V

Plasmid extraction

by Pauline

Biobricks:

  • BBa_K1707003 #4 et #5

With Plasmid DNA purification kit from Macherey-Nagel

Digestion + Purification on agarose gel

by Pauline

Biobricks:

  • BBa_K1707003 #4 and #5

Mix:

  • 10µL plasmid BBa_K1707003
  • 1µL XbaI
  • 1µL PstI
  • 2µL Buffer FastDigest
  • 6µL H2O

Purification on agarose gel 1%, migration 100V

ParisSaclay 27.07.15-quantification+vérif.jpg

Wells 2-3: Quantification, Wells 9-10: Verification of purification; from left to right: 1. DNA Ladder, 2. BBa_K1707000#2, 3. BBa_50051, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. BBa_K1707003#1, 10. BBa_K1707003#2, 11. Empty 12. Empty

Cut with a scalpel Purification with the PCR Clean up kit from Macherey-Nagel

New Culture

by Johan

Strains:

  • 1696
  • 1693
  • 1320

in 150mL LB without any antibiotic




Member present:

  • Instructors: Alice
  • Students: Coralie, Audrey, Pauline, Johan and Seong-Koo

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Contact
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France Copyright © 2015 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.