Difference between revisions of "Team:Paris Saclay/Notebook/July/22"

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=Wednesday 22nd July=
 
=Wednesday 22nd July=
 
==Lab Work==
 
==Lab Work==
−
===Titre de l'Experience===
+
===Electrophoresis===
−
''by coffee boy''
+
''by Johan and Pauline''
 +
* PCR Products:
 +
** BBa_J23101
 +
** BBa_K1707000
 +
* Purification quantification:
 +
** BBa_K1707002
 +
** BBa_J23101
 +
** BBa_R0051
 +
** BBa_K098997
 +
** BBa_K1707001
 +
* Undigested plasmids:
 +
** BBa_K1707001 #1 and #2
 +
** BBa_K1707002 #1 and #2
 +
** BBa_K1707003 #2
 +
** BBa_J23101
 +
** BBa_B0015
 +
We can't see PCR products. We can suppose that they run out the gel.
 +
[[File:Paris Saclay-22.07.2015 - Plasmides 2.jpg|300px|center]]
 +
<html><i><p>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707001#1, 3. BBa_K1707001#2, 4. BBa_K1707002#1, 5. BBa_K1707002#2, 6. BBa_K1707003#2, 7. BBa_J23101, 8. BBa_B0015, 9. Empty, 10. Empty, 11. Empty, 12. Empty</p></i></html>
 +
[[File:Paris Saclay-22.07.15 - Quantification 1.jpg|300px|center]]
 +
<html><i><p>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. PCR J23101, 3. PCR K1707000#1, 4. PCR K1707000#2, 5. BBa_K1707002, 6. BBa_J23101, 7. BBa_R0051, 8. BBa_K098997, 9. BBa_K1707001, 10. Empty</p></i></html>
  
−
Voilà, ça c'est le standard pour le notebook. Pour modifier cette page, allez au ménou '''wiki tools''' en haut à gauche, puis sur '''Edit this page'''. Si vous avez des questions, n'hésitez pas à me contacter. ''Bien évidemment ce text doit être supprimer une fois le notebook rempli :) Bonnes manips!
+
===Digestion===
 +
'' by Coralie''
 +
 
 +
*Digestion by EcoRI
 +
**Mix:
 +
*** 7µL EcoRI
 +
*** 7µL Buffer FastDigest 10x
 +
*** 42µL H2O
 +
8µL in each tube + 2µL Plamid
 +
** Biobricks:
 +
*** BBa_B0015
 +
*** BBa_J23101
 +
*** BBa_K1707003 #2
 +
*** BBa_K1707002 #1 and #2
 +
*** BBa_K1707001 #1 and #2
 +
 
 +
*Digestion by EcoRI + PstI
 +
**Mix:
 +
*** 7µL EcoRI
 +
*** 7µL PstI
 +
*** 7µL Buffer FastDigest 10x
 +
*** 35µL H2O
 +
8µL in each tube + 2µL Plamid
 +
** Biobricks:
 +
*** BBa_B0015
 +
*** BBa_J23101
 +
*** BBa_K1707003 #2
 +
*** BBa_K1707002 #1 and #2
 +
*** BBa_K1707001 #1 and #2
 +
 
 +
Incubation 2h, 37°C
 +
 
 +
===Electrophoresis===
 +
''by Johan''
 +
[[File:Paris Saclay-22.07.2015 - Gel vérif simple digestion EcoRI.jpg|300px|center]]
 +
<html><p><i>Verification by digestion with EcoRI, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty</i></p></html>
 +
[[File:Paris Saclay-22.07.15 - Vérification double digestion EcoRI + PstI.jpg|300px|center]]
 +
<html><p><i>Verification by digestion with EcoRI and PstI, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty, 11. Empty, 12. Empty</i></p></html>
 +
We can conclude that K1707003 #2 isn't OK
 +
 
 +
===Plasmid extraction===
 +
''by Pauline and Seong Koo''
 +
 
 +
Biobricks:
 +
* BBa_K1707003 #4 to #9
 +
* BBa_C0051
 +
* BBa_E0240
 +
* BBa_J06702
 +
 
 +
 
 +
===Soil experiment===
 +
''by Audrey and Johan''
 +
 
 +
Plates observation and count of CFU:
 +
* Strain 1696:
 +
** Dilution 1: more than 1000
 +
** Dilution 0.1: more than 1000
 +
** Dilution 0.01: 441 CFU
 +
 
 +
* Strain 1693:
 +
** Dilution 1: more than 1000
 +
** Dilution 0.1: more than 1000
 +
** Dilution 0.01: 414 CFU
 +
 
 +
* Strain 1320:
 +
** Dilution 1: more than 1000
 +
** Dilution 0.1: 1744 CFU
 +
** Dilution 0.01: 272 CFU
 +
 
 +
We take 1g of contaminated soil and treat it like yesterday to spread it on specific plates
 +
Incubation ON, 37°C
 +
 
 +
===Electrophoresis of PCR products===
 +
 
 +
[[File:ParisSaclay 22.07.15 ARN tige boucle PCR.jpg|300px|center]]
 +
<html><p><i>Verification PCR products, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. Indicator (BBa_J23101), 3. BBa_K1707000#1, 4. BBa_K1707000#2</i></p></html>
  
 
'''Member present:'''
 
'''Member present:'''
−
* Instructors:
+
* Instructors: Alice
−
* Students:  
+
* Students: Johan, Seong Koo, Pauline, Audrey, Coralie
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 21:17, 18 September 2015

Wednesday 22nd July

Lab Work

Electrophoresis

by Johan and Pauline

  • PCR Products:
    • BBa_J23101
    • BBa_K1707000
  • Purification quantification:
    • BBa_K1707002
    • BBa_J23101
    • BBa_R0051
    • BBa_K098997
    • BBa_K1707001
  • Undigested plasmids:
    • BBa_K1707001 #1 and #2
    • BBa_K1707002 #1 and #2
    • BBa_K1707003 #2
    • BBa_J23101
    • BBa_B0015

We can't see PCR products. We can suppose that they run out the gel.

Paris Saclay-22.07.2015 - Plasmides 2.jpg

Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707001#1, 3. BBa_K1707001#2, 4. BBa_K1707002#1, 5. BBa_K1707002#2, 6. BBa_K1707003#2, 7. BBa_J23101, 8. BBa_B0015, 9. Empty, 10. Empty, 11. Empty, 12. Empty

Paris Saclay-22.07.15 - Quantification 1.jpg

Quantification, from left to right: 1. DNA Ladder, 2. PCR J23101, 3. PCR K1707000#1, 4. PCR K1707000#2, 5. BBa_K1707002, 6. BBa_J23101, 7. BBa_R0051, 8. BBa_K098997, 9. BBa_K1707001, 10. Empty

Digestion

by Coralie

  • Digestion by EcoRI
    • Mix:
      • 7µL EcoRI
      • 7µL Buffer FastDigest 10x
      • 42µL H2O

8µL in each tube + 2µL Plamid

    • Biobricks:
      • BBa_B0015
      • BBa_J23101
      • BBa_K1707003 #2
      • BBa_K1707002 #1 and #2
      • BBa_K1707001 #1 and #2
  • Digestion by EcoRI + PstI
    • Mix:
      • 7µL EcoRI
      • 7µL PstI
      • 7µL Buffer FastDigest 10x
      • 35µL H2O

8µL in each tube + 2µL Plamid

    • Biobricks:
      • BBa_B0015
      • BBa_J23101
      • BBa_K1707003 #2
      • BBa_K1707002 #1 and #2
      • BBa_K1707001 #1 and #2

Incubation 2h, 37°C

Electrophoresis

by Johan

Paris Saclay-22.07.2015 - Gel vérif simple digestion EcoRI.jpg

Verification by digestion with EcoRI, from left to right: 1. DNA Ladder, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty

Paris Saclay-22.07.15 - Vérification double digestion EcoRI + PstI.jpg

Verification by digestion with EcoRI and PstI, from left to right: 1. DNA Ladder, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty, 11. Empty, 12. Empty

We can conclude that K1707003 #2 isn't OK

Plasmid extraction

by Pauline and Seong Koo

Biobricks:

  • BBa_K1707003 #4 to #9
  • BBa_C0051
  • BBa_E0240
  • BBa_J06702


Soil experiment

by Audrey and Johan

Plates observation and count of CFU:

  • Strain 1696:
    • Dilution 1: more than 1000
    • Dilution 0.1: more than 1000
    • Dilution 0.01: 441 CFU
  • Strain 1693:
    • Dilution 1: more than 1000
    • Dilution 0.1: more than 1000
    • Dilution 0.01: 414 CFU
  • Strain 1320:
    • Dilution 1: more than 1000
    • Dilution 0.1: 1744 CFU
    • Dilution 0.01: 272 CFU

We take 1g of contaminated soil and treat it like yesterday to spread it on specific plates Incubation ON, 37°C

Electrophoresis of PCR products

ParisSaclay 22.07.15 ARN tige boucle PCR.jpg

Verification PCR products, from left to right: 1. DNA Ladder, 2. Indicator (BBa_J23101), 3. BBa_K1707000#1, 4. BBa_K1707000#2

Member present:

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

Back to the calendar

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.