Difference between revisions of "Team:Paris Saclay/Notebook/August/19"

(→‎Colony and Extraction product DreamTaq PCR)
(→‎Plasmids extraction)
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!260/280
 
!260/280
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_009 (GFP1-9) clone 4
−
|
+
|207.03
−
|
+
|1.76
−
|
+
|1.85
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_009 (GFP1-9) clone 5
−
|
+
|280.30
−
|
+
|1.79
−
|
+
|1.89
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_014 (sgRNA Nm) clone 1
−
|
+
|350.24
−
|
+
|2.24
−
|
+
|1.97
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_014 (sgRNA Nm) clone 2
−
|
+
|187.62
−
|
+
|2.20
−
|
+
|1.05
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_013 (FKBP/pJET)clone 3
−
|
+
|272.37
−
|
+
|2.32
−
|
+
|1.93
 
|-
 
|-
−
|pPS16_0XX clone X
+
|pPS16_013 (FKBP/pJET)clone 4
−
|
+
|404.65
−
|
+
|2.34
−
|
+
|1.94
 +
|-
 +
|pPS16_013 (FKBP/pJET)clone 5
 +
|204.87
 +
|2.32
 +
|1.94
 +
|-
 +
|pPS16_010 (FRB) clone 2
 +
|256.44
 +
|2.35
 +
|1.96
 +
|-
 +
|pPS16_010 (FRB) clone 3
 +
|73.04
 +
|2.13
 +
|2.05
 +
|-
 +
|pPS16_010 (FRB) clone 5
 +
|129.86
 +
|2.16
 +
|1.87
 +
|-
 +
|pPS16_002 (1.2) clone 1
 +
|49.72
 +
|1.34
 +
|1.67
 +
|-
 +
|pPS16_002 (1.2) clone 11
 +
|392.66
 +
|2.33
 +
|1.95
 
|-
 
|-
 
|}
 
|}

Revision as of 10:35, 19 August 2016

Friday 19th August

Lab work

Visualization

Plasmids extraction

"By Terrence, Alice and Manhaz"

Nano drop :

Plasmid name Concentration (ng/µL) 260/230 260/280
pPS16_009 (GFP1-9) clone 4 207.03 1.76 1.85
pPS16_009 (GFP1-9) clone 5 280.30 1.79 1.89
pPS16_014 (sgRNA Nm) clone 1 350.24 2.24 1.97
pPS16_014 (sgRNA Nm) clone 2 187.62 2.20 1.05
pPS16_013 (FKBP/pJET)clone 3 272.37 2.32 1.93
pPS16_013 (FKBP/pJET)clone 4 404.65 2.34 1.94
pPS16_013 (FKBP/pJET)clone 5 204.87 2.32 1.94
pPS16_010 (FRB) clone 2 256.44 2.35 1.96
pPS16_010 (FRB) clone 3 73.04 2.13 2.05
pPS16_010 (FRB) clone 5 129.86 2.16 1.87
pPS16_002 (1.2) clone 1 49.72 1.34 1.67
pPS16_002 (1.2) clone 11 392.66 2.33 1.95

Colony and Extraction product DreamTaq PCR

"By Léa and Naiane"

A Colony PCR was performed on Dh5a transformed with PJet containing sgRNA Nm or 1.2 gBlocks. sgRNA Nm clones 6, 7, 8, 10, 11 and 12 were screened, and 1.2 clones 13 to 18 were screened and plated on Petri dish containing solid LB and Ampicilin.

In the meantime, a DreamTaq PCR was performed on plasmids extractions listed above, using the same PCR mix. The PCR mix was made following the usual protocol. Universal primers for PJET were used (Tm= 51.9°).

A previous step of denaturation (95°C, 5min) was performed for colony PCR.





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