Team:Paris Saclay/Notebook/July/30

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==Lab Work==
==Lab Work==
 +
 +
===C - Lemon scent===
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 +
====PCR of BBa_K762100====
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''by Sean''
 +
 +
Oligonucleotides used: iPS66, iPS67
 +
 +
Oligonucleotides were diluted twice from 100µM to 50µM.
 +
 +
'''Protocol'''
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 +
Add into a PCR tube the following:
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
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! scope=col | Component
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! scope=col | For a total volume of 50μl
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|-
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|H<sub>2</sub>O
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|35.5μl
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|-
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| Phusion buffer 5X
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| 10μl
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|-
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| dNTPs
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| 1μl
 +
|-
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| iPS66
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| 1μl
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|-
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| iPS67
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| 1μl
 +
|-
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| BBa_K762100
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| 1μl
 +
|-
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| Phusion DNA polymerase
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| 0.25μl
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|}
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 +
Tube was 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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|-
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| width="25%" |
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Initial denaturation
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| width="25%" |
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98°C
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| width="25%" |
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1 min
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| width="25%" |
 +
1
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|-
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| Denaturation
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| 98°C
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| 15 s
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| 25 - 30
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|-
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| Annealing
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| variable
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| 55°C
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| 25 - 30
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|-
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| Extension
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| 72°C
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| 45 s
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| 25-30
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|-
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| Final extension
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| 72°C
 +
| 10 min
 +
| 1
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|-
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| Final extension
 +
| 8°C
 +
| hold
 +
| 1
 +
|}

Revision as of 14:48, 30 July 2014

Contents

Wednesday 30th July

Lab Work

C - Lemon scent

PCR of BBa_K762100

by Sean

Oligonucleotides used: iPS66, iPS67

Oligonucleotides were diluted twice from 100µM to 50µM.

Protocol

Add into a PCR tube the following:

Component For a total volume of 50μl
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 1μl
iPS66 1μl
iPS67 1μl
BBa_K762100 1μl
Phusion DNA polymerase 0.25μl

Tube was 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 variable 55°C 25 - 30
Extension 72°C 45 s 25-30
Final extension 72°C 10 min 1
Final extension 8°C hold 1
Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/July/30"