Team:Paris Saclay/Notebook/August/7

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===D - Lemon Sent===
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 +
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==== Polymerase chain reaction ====
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''by Sean & Pierre''
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 +
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for this PCR, five tubes of each of the following Bio-bricks were prepared.
 +
 +
'''BBa_K517003'''
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
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! scope=col | component
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! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
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|35.5μl
 +
|-
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| Phusion buffer 5X
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| 10μl
 +
|-
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| dNTPs
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| 2μl
 +
|-
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| iPS68bis
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| 1μl
 +
|-
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| iPS69
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| 1μl
 +
|-
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| DNA
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| 1μl
 +
|-
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| Phusion enzyme
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| 0.5μl
 +
|}
 +
 +
 +
'''BBa_K762100'''
 +
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|35.5μl
 +
|-
 +
|Phusion buffer 5X
 +
|10μl
 +
|-
 +
|dNTPs
 +
|2μl
 +
|-
 +
|iPS66
 +
|1μl
 +
|-
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|iPS67
 +
|1μl
 +
|-
 +
|DNA
 +
|1μl
 +
|-
 +
|Phusion enzyme
 +
|0.5μl
 +
|}
 +
 +
Tubes were placed in PCR machine with the following parameters.
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 +
{| class="wikitable centre" width="80%"
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|+
 +
|-
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! scope=col | Cycle step
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! scope=col | Temperature
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! scope=col | Time
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! scope=col | Cycle
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|-
 +
| width="25%" |
 +
Initial denaturation
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| width="25%" |
 +
98°C
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| width="25%" |
 +
1 min
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| width="25%" |
 +
1
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|-
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| Denaturation
 +
| 98°C
 +
| 15 s
 +
| 25 - 30
 +
|-
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| Annealing
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| 52°C
 +
| 25 s
 +
| 25 - 30
 +
|-
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| Extension
 +
| 72°C
 +
| 45 s
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| 25-30
 +
|-
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| Final extension
 +
| 72°C
 +
| 10 min
 +
| 1
 +
|-
 +
| Final extension
 +
| 8°C
 +
| hold
 +
| 1
 +
|}

Revision as of 13:50, 7 August 2014

Contents

Thursday 7th August

Lab work

C - Salicylate Inducible Suppressing System

DNA extraction from gel agarose

In progress


Members present:

  • Instructors and advisors: Alice.
  • Students: Eugene, Fabio, Hoang Vu, Juliette, Melanie, Pierre, Romain, Sean and Terry.

Back to the calendar


D - Lemon Sent

Polymerase chain reaction

by Sean & Pierre


for this PCR, five tubes of each of the following Bio-bricks were prepared.

BBa_K517003

component volume
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 2μl
iPS68bis 1μl
iPS69 1μl
DNA 1μl
Phusion enzyme 0.5μl


BBa_K762100


component volume
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 2μl
iPS66 1μl
iPS67 1μl
DNA 1μl
Phusion enzyme 0.5μl

Tubes were placed in PCR machine with the following parameters.

Cycle step Temperature Time Cycle

Initial denaturation

98°C

1 min

1

Denaturation 98°C 15 s 25 - 30
Annealing 52°C 25 s 25 - 30
Extension 72°C 45 s 25-30
Final extension 72°C 10 min 1
Final extension 8°C hold 1
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