Difference between revisions of "Team:Paris Saclay/Notebook/August/3"

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(→‎Extraction of the plasmids containing gBlocks pPS16_001, pPS16_002, pPS16_003, FRB, sg-ST1, FKPB, DS-NMcasN-, spacer and detection)
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The extraction was performed without kit, following the usual [[Team:Paris_Saclay/Experiments#Plasmid_DNA_extraction|protocol]].
 
The extraction was performed without kit, following the usual [[Team:Paris_Saclay/Experiments#Plasmid_DNA_extraction|protocol]].
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It was done on:
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{| class="wikitable"
 +
|-
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! Puc 19 containing
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! Clone
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|-
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| pPS16_001
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| 4
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|-
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| pPS16_002
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| 3
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|-
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| pPS16_003
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| 2, 5
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|-
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| FRB
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| 5
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|-
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| sg-ST1
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| 2, 3, 5
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|-
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| DS-NMcasN-
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| 7, 11
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|-
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| FKBP
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| 4
 +
|-
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| Spacer
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| 1, 6
 +
|-
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| Detection
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| 3, 4, 6
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|}
  
 
====Extraction of the plasmids containing gBlocks====
 
====Extraction of the plasmids containing gBlocks====

Revision as of 14:03, 4 August 2016

Tuesday 3st August

Lab work

Visualization

Glycerol stocks

By Caroline

The bacteria transformed with the plasmids sent the 2/08/2016 and those that would be sent the 4/08/2016 were put into glycerol. 1mL of liquid culture and 0.5mL of glycerol were put at -20°C.

High fidelity Q5 PCR on DH5alp|pPS16_004, DH5alp|pPS16_006 and DH5alp|pPS16_007

By Caroline

Q5 PCR was carried out following the usual protocol adapted to 50µL at a TM of 72°C. The primers used were specific to amplify only the interested sequence.

Extraction of the plasmids containing gBlocks pPS16_001, pPS16_002, pPS16_003, FRB, sg-ST1, FKPB, DS-NMcasN-, spacer and detection

By Laetitia

The extraction was performed without kit, following the usual protocol.

It was done on:

Puc 19 containing Clone
pPS16_001 4
pPS16_002 3
pPS16_003 2, 5
FRB 5
sg-ST1 2, 3, 5
DS-NMcasN- 7, 11
FKBP 4
Spacer 1, 6
Detection 3, 4, 6

Extraction of the plasmids containing gBlocks

By Naiane

The kit "Charge Switch-Pro Plasmid Miniprep" was used to extract plasmidic DNA of the plasmid pPS16_003 containing the Gblock 2.1, plasmid pPS16_004 containing the Gblock 2.2, plasmid pPS16_006 containing the Gblock 3.2 and the plasmid pPS16_007 containing the Gblock 4.1 from 3mL of overnight culture. Plasmids were resuspended in 100μL of Milli-Q water. DNA stored at -20°C.

Interlab Study

By






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