Difference between revisions of "Team:Paris Saclay/Notebook/August/20"

(Created page with "=Thursday 20th August= ==Lab Work== ===Digestion for check=== ''by Audrey'' Biobricks: * BBa_K115017 (control +) * BBa_J23101 + GFP (control -) * BBa_K1707000 #2, #3 and #4 * ...")
 
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(One intermediate revision by one other user not shown)
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{{Team:Paris_Saclay/notebook_header}}
 
=Thursday 20th August=
 
=Thursday 20th August=
 
==Lab Work==
 
==Lab Work==
−
 
  
 
===Digestion for check===
 
===Digestion for check===
 
''by Audrey''
 
''by Audrey''
  
−
Biobricks:
 
 
* BBa_K115017 (control +)
 
* BBa_K115017 (control +)
 
* BBa_J23101 + GFP (control -)
 
* BBa_J23101 + GFP (control -)
Line 25: Line 24:
 
** 2 µL Buffer 3 10x (Biolabs)
 
** 2 µL Buffer 3 10x (Biolabs)
 
** 5 µL H2O
 
** 5 µL H2O
−
 
  
 
===Electrophoresis to check PCR products===
 
===Electrophoresis to check PCR products===
Line 34: Line 32:
 
Biobricks:
 
Biobricks:
 
* BBa_K1707031 #7 to #26
 
* BBa_K1707031 #7 to #26
−
 
+
[[File:ParisSaclay 20.08.15-PCR1.jpg|300px|center]]
 +
<html><p><i>Verification of BBa_K1707031 PCR products, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. #7, 3. #8, 4. #9, 5. #10, 6. #11, 7. #12, 8, #13, 9. #14, 10. #15, 11. #16, 12. #17</i></p></html>
 +
[[File:ParisSaclay 20.08.15-PCR2.jpg|300px|center]]
 +
<html><p><i>Verification of BBa_K1707031 PCR products, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. #18, 3. #19, 4. #20, 5. #21, 6. #22, 7. #23, 8, #24, 9. #25, 10. #26</i></p></html>
 
We can conclude that all clones aren't OK
 
We can conclude that all clones aren't OK
−
 
  
 
===Purification===
 
===Purification===
 
''by Pauline''
 
''by Pauline''
  
−
Biobricks:
 
 
* BBa_K1707022 #1
 
* BBa_K1707022 #1
 
* BBa_K1707023 #1
 
* BBa_K1707023 #1
Line 55: Line 54:
  
 
Digestion products:
 
Digestion products:
−
Biobricks:
 
 
* BBa_K115017 (control +)
 
* BBa_K115017 (control +)
 
* BBa_J23101 + GFP (control -)
 
* BBa_J23101 + GFP (control -)
Line 67: Line 65:
 
* BBa_K1707035 #1
 
* BBa_K1707035 #1
 
* BBa_K1707036 #1
 
* BBa_K1707036 #1
−
 
+
[[File:ParisSaclay 20.08.15-restriction1.jpg|300px|center]]
 +
<html><p><i>Verification by digestion with SfoI and BspEI, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K115017, 3. BBa_K1707039, 4. BBa_K1707000#2, 5. BBa_K1707000#3, 6. BBa_K1707000#4, 7. BBa_K1707013#1, 8. BBa_K1707030#1, 9. BBa_K1707019#1, 10. BBa_K1707020#2, 11. BBa_K1707021#2, 12. BBa_K1707027#1</i></p></html>
 +
[[File:ParisSaclay 20.08.15-restriction2+quantif.jpg|300px|center]]
 +
<html><p><i>Wells 1-2: Verification by digestion with SfoI and BspEI, Wells 4-8: Quantification; from left to right: 1. BBa_K1707035#1, 2. BBa_K1707036#1, 3. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 4. BBa_K1707004#2, 5. BBa_K1707022#1, 6. BBa_K1707023#1, 7. BBa_K1707023#1, 8. BBa_K1707034#1, 9. Empty, 10. Empty</i></p></html>
 
Quantification:
 
Quantification:
−
Biobricks:
 
 
* BBa_K1707022 #1: ?
 
* BBa_K1707022 #1: ?
 
* BBa_K1707023 #1: nothing
 
* BBa_K1707023 #1: nothing
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''by Pauline''
 
''by Pauline''
  
−
Biobrick: BBa_K1707004 #2
+
* BBa_K1707004 #2
  
 
In 5mL LB + 5µL Ampicillin
 
In 5mL LB + 5µL Ampicillin
−
 
−
 
  
 
===Ligation===
 
===Ligation===
 
''by Pauline''
 
''by Pauline''
  
−
BBa_K1707037= BBa_K1707028 + BBa_K1707034
+
* BBa_K1707037 = BBa_K1707028 + BBa_K1707034
  
 
Mix:
 
Mix:
Line 99: Line 97:
  
 
Incubation 4°C, ON
 
Incubation 4°C, ON
−
 
  
 
===Quantification===
 
===Quantification===
Line 107: Line 104:
 
* BBa_K1707022 #1
 
* BBa_K1707022 #1
 
* BBa_K1707023 #1
 
* BBa_K1707023 #1
−
 
+
[[File:ParisSaclay 20.08.15-quantif2.jpg|300px|center]]
 +
<html><p><i>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707022#1, 3. BBa_K1707023#1, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty</i></p></html>
 
We can't see anything
 
We can't see anything
−
 
−
 
−
 
  
 
'''Member present:'''
 
'''Member present:'''

Latest revision as of 22:55, 18 September 2015

Thursday 20th August

Lab Work

Digestion for check

by Audrey

  • BBa_K115017 (control +)
  • BBa_J23101 + GFP (control -)
  • BBa_K1707000 #2, #3 and #4
  • BBa_K1707013 #1
  • BBa_K1707030 #1
  • BBa_K1707019 #1
  • BBa_K1707020 #2
  • BBa_K1707021 #2
  • BBa_K1707027 #1
  • BBa_K1707035 #1
  • BBa_K1707036 #1

Mix for each tub:

    • 2 µL plasmid
    • 1 µL SpeI (Biolabs)
    • 1 µL BspE1 (Biolabs)
    • 2 µL Buffer 3 10x (Biolabs)
    • 5 µL H2O

Electrophoresis to check PCR products

by Audrey

Agarose gel 1%, Migration 90V

Biobricks:

  • BBa_K1707031 #7 to #26
ParisSaclay 20.08.15-PCR1.jpg

Verification of BBa_K1707031 PCR products, from left to right: 1. DNA Ladder, 2. #7, 3. #8, 4. #9, 5. #10, 6. #11, 7. #12, 8, #13, 9. #14, 10. #15, 11. #16, 12. #17

ParisSaclay 20.08.15-PCR2.jpg

Verification of BBa_K1707031 PCR products, from left to right: 1. DNA Ladder, 2. #18, 3. #19, 4. #20, 5. #21, 6. #22, 7. #23, 8, #24, 9. #25, 10. #26

We can conclude that all clones aren't OK

Purification

by Pauline

  • BBa_K1707022 #1
  • BBa_K1707023 #1
  • BBa_K1707004 #2
  • BBa_K1707034 #1
  • BBa_K1707028 #1

With Macherey Nagel purification kit


Electrophoresis

by Pauline

Digestion products:

  • BBa_K115017 (control +)
  • BBa_J23101 + GFP (control -)
  • BBa_K1707000 #2, #3 and #4
  • BBa_K1707013 #1
  • BBa_K1707030 #1
  • BBa_K1707019 #1
  • BBa_K1707020 #2
  • BBa_K1707021 #2
  • BBa_K1707027 #1
  • BBa_K1707035 #1
  • BBa_K1707036 #1
ParisSaclay 20.08.15-restriction1.jpg

Verification by digestion with SfoI and BspEI, from left to right: 1. DNA Ladder, 2. BBa_K115017, 3. BBa_K1707039, 4. BBa_K1707000#2, 5. BBa_K1707000#3, 6. BBa_K1707000#4, 7. BBa_K1707013#1, 8. BBa_K1707030#1, 9. BBa_K1707019#1, 10. BBa_K1707020#2, 11. BBa_K1707021#2, 12. BBa_K1707027#1

ParisSaclay 20.08.15-restriction2+quantif.jpg

Wells 1-2: Verification by digestion with SfoI and BspEI, Wells 4-8: Quantification; from left to right: 1. BBa_K1707035#1, 2. BBa_K1707036#1, 3. DNA Ladder, 4. BBa_K1707004#2, 5. BBa_K1707022#1, 6. BBa_K1707023#1, 7. BBa_K1707023#1, 8. BBa_K1707034#1, 9. Empty, 10. Empty

Quantification:

  • BBa_K1707022 #1: ?
  • BBa_K1707023 #1: nothing
  • BBa_K1707004 #2: nothing
  • BBa_K1707034 #1: 20 µg/µL
  • BBa_K1707028 #1: 10 µg/µL


New culture

by Pauline

  • BBa_K1707004 #2

In 5mL LB + 5µL Ampicillin

Ligation

by Pauline

  • BBa_K1707037 = BBa_K1707028 + BBa_K1707034

Mix:

  • 8 µL BBa_K1707028
  • 10 µL BBa_K1707034
  • 2 µL Ligase
  • 2,5 µL Buffer
  • 2,5 µL H2O

Incubation 4°C, ON

Quantification

by Pauline

BIobricks:

  • BBa_K1707022 #1
  • BBa_K1707023 #1
ParisSaclay 20.08.15-quantif2.jpg

Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707022#1, 3. BBa_K1707023#1, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty

We can't see anything

Member present:

  • Instructors: Claire
  • Students: Pauline and Audrey

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.