Difference between revisions of "Team:Paris Saclay/Notebook/July/31"

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(Electrophoresis)
 
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Migration 100V
 
Migration 100V
 
[[File:Paris Saclay-31.07.2015 - Verification K012-013.jpg|300px|center]]
 
[[File:Paris Saclay-31.07.2015 - Verification K012-013.jpg|300px|center]]
<html><i><p>Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. DNA Ladder, 3. BBa_K1707012#1, 4. BBa_K1707012#2, 5. BBa_K1707012#3, 6. BBa_K1707012#4, 7. BBa_K1707013#1, 8. BBa_K1707013#2, 9. BBa_K1707013#3, 10. BBa_K1707013#2, 11. Empty, 12. Empty</p></i></html>
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<html><i><p>Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 3. BBa_K1707012#1, 4. BBa_K1707012#2, 5. BBa_K1707012#3, 6. BBa_K1707012#4, 7. BBa_K1707013#1, 8. BBa_K1707013#2, 9. BBa_K1707013#3, 10. BBa_K1707013#2, 11. Empty, 12. Empty</p></i></html>
 
[[File:Paris Saclay-31.07.2015 - Vérification K019.jpg|300px|center]]
 
[[File:Paris Saclay-31.07.2015 - Vérification K019.jpg|300px|center]]
<html><i><p>Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. DNA Ladder, 3. BBa_K1707019#1, 4. BBa_K1707019#2, 5. BBa_K1707019#3, 6. BBa_K1707019#4, 7. Empty</p></i></html>
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<html><i><p>Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 3. BBa_K1707019#1, 4. BBa_K1707019#2, 5. BBa_K1707019#3, 6. BBa_K1707019#4, 7. Empty</p></i></html>
 
We can conclude that all biobricks are OK
 
We can conclude that all biobricks are OK
  

Latest revision as of 21:26, 18 September 2015


Friday 31st July

Lab Work

Plasmid extraction

by Seong-Koo

Biobricks:

  • BBa_K1707012 #1, #2, #3 and #4
  • BBa_K1707013 #1, #2, #3 and #4
  • BBa_K1707019 #1, #2, #3 and #4

With the Macherey-Nagel Extraction kit


Rehydratation

by Pauline

Biobrick: BBa_I13600 (2014-3-19C and 2013-3-20C)


Transformation

by Pauline

Biobricks:

  • BBa_I13600 (2013, 2014 and 2015)
  • BBa_K1707008

As usual

Plasmid extraction

by Pauline

Biobricks:

  • BBa_E0422 #1 and #2
  • BBa_E0022 #1 and #2

With the Macherey-Nagel Extraction kit

New culture

by Pauline

Biobricks:

  • BBa_K1707004: amicillin
  • BBa_K1707011
  • BBa_K1707021

6 clones of each in 5 mL LB

Digestion Verification

by Coralie

  • Biobricks:
    • BBa_K1707012 #1, #2, #3 and #4
    • BBa_K1707013 #1, #2, #3 and #4
    • BBa_K1707019 #1, #2, #3 and #4
  • Mix:
    • 1 µL Buffer FastDigest 10x
    • 0,5 µL XbaI
    • 0,5 µL PstI
    • 2 µL Plasmid
    • 6 µL H2O

Incubation 2h, 37°C

Electrophoresis

by Coralie

Agarose gel, 1% Migration 100V

Paris Saclay-31.07.2015 - Verification K012-013.jpg

Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. DNA Ladder, 3. BBa_K1707012#1, 4. BBa_K1707012#2, 5. BBa_K1707012#3, 6. BBa_K1707012#4, 7. BBa_K1707013#1, 8. BBa_K1707013#2, 9. BBa_K1707013#3, 10. BBa_K1707013#2, 11. Empty, 12. Empty

Paris Saclay-31.07.2015 - Vérification K019.jpg

Verification by digestion with XbaI and PstI, from left to right: 1. Empty, 2. DNA Ladder, 3. BBa_K1707019#1, 4. BBa_K1707019#2, 5. BBa_K1707019#3, 6. BBa_K1707019#4, 7. Empty

We can conclude that all biobricks are OK

Soil experiment

by Coralie and Johan

Soil

Observation of J0 plates: there are a lot of colony in each.

Control (-):

  • MCK + Tet: 40 pink colonies
  • MCK + Cm: 10 pink colonies
  • MCK + Spectino: 1 pink colony
  • MCK: 100 pink colonies and a lot of white colonies

We can suppose that we have a contamination between our experiment and control. Or maybe we have a MCK problem because we change it during the experiment.

So, we decide to stop the experiment and do some test:

  • Soil + H2O on new MCK
  • Soil + LB on new MCK
  • LB on new MCK
  • MCK alone

Incubation over night, 37°C


New culture

by Pauline

  • BBa_K1707005
  • BBa_I13602

With plasmid: PIJ773

Incubation at 37°C, 250 rpm

Member present:

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

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