Team:Paris Saclay/Notebook/August/27

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(Difference between revisions)
(→Plasmide extraction)
(→PCR of Limonene synthase (BBa762100)=)
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Extraction of pPS5 with the phenol protocol (as previously described)
Extraction of pPS5 with the phenol protocol (as previously described)
-
====PCR of Limonene synthase (BBa762100)=====
+
====PCR of Limonene synthase (BBa762100)====
 +
''by Mélanie''
 +
 
 +
5 tubes to do a gradient
 +
 
 +
{| class="wikitable centre" width="50%"
 +
|+ 
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|H<sub>2</sub>O
 +
|29.75μl
 +
|-
 +
|Gotaq buffer
 +
|10µl
 +
|-
 +
|dNTP 10mM
 +
|1µl
 +
|-
 +
|iPS67
 +
|1µM
 +
|-
 +
|iPS66
 +
|1µM
 +
|-
 +
|DNA
 +
|1µl
 +
|-
 +
|MgCl2
 +
|4µl
 +
|-
 +
|Gotaq
 +
|1.25µl
 +
|}
 +
 
 +
{| class="wikitable centre" width="50%"
 +
|+ PCR cycle
 +
|-
 +
! scope=col | step
 +
! scope=col | temperature (°C)
 +
! scope=col | time
 +
|-
 +
|1
 +
|95
 +
|2min
 +
|-
 +
|2
 +
|95
 +
|30 sec
 +
|-
 +
|3
 +
|49-55 (gradient)
 +
|30sec
 +
|-
 +
|4
 +
|72
 +
|2min
 +
|-
 +
|5
 +
|72
 +
|5min
 +
|}
 +
 
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 14:42, 27 August 2014

Contents

Wednesday 27th August

Lab Work

D - lemon scent

Plasmide extraction

By Mélanie

Extraction of pPS5 with the phenol protocol (as previously described)

PCR of Limonene synthase (BBa762100)

by Mélanie

5 tubes to do a gradient

components volumes
H2O 29.75μl
Gotaq buffer 10µl
dNTP 10mM 1µl
iPS67 1µM
iPS66 1µM
DNA 1µl
MgCl2 4µl
Gotaq 1.25µl
PCR cycle
step temperature (°C) time
1 95 2min
2 95 30 sec
3 49-55 (gradient) 30sec
4 72 2min
5 72 5min


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