Line 6:
Line 6: ====Plasmids extraction====
====Plasmids extraction====
"By Terrence, Alice and Mahnaz"
"By Terrence, Alice and Mahnaz"
+
+ The following plasmids were extracted using the [[Team:Paris_Saclay/Experiments#InvitrogenPlasmidExtraction|"Charge Switch-Pro Plasmid Miniprep"]] kit:
+ *pPS16_009 (GFP1-9) clone 4
+ *pPS16_009 (GFP1-9) clone 5
+ *pPS16_014 (sgRNA Nm) clone 1
+ *pPS16_014 (sgRNA Nm) clone 2
+ *pPS16_013 (FKBP/pJET) clone 3
+ *pPS16_013 (FKBP/pJET) clone 4
+ *pPS16_013 (FKBP/pJET) clone 5
+ *pPS16_010 (FRB) clone 2
+ *pPS16_010 (FRB) clone 3
+ *pPS16_010 (FRB) clone 5
+ *pPS16_002 (1.2) clone 1
+ *pPS16_002 (1.2) clone 11
+
Nano drop :
Nano drop :
Revision as of 13:59, 19 August 2016
Friday 19th August Lab work Visualization "By Terrence, Alice and Mahnaz"
The following plasmids were extracted using the "Charge Switch-Pro Plasmid Miniprep" kit:
pPS16_009 (GFP1-9) clone 4
pPS16_009 (GFP1-9) clone 5
pPS16_014 (sgRNA Nm) clone 1
pPS16_014 (sgRNA Nm) clone 2
pPS16_013 (FKBP/pJET) clone 3
pPS16_013 (FKBP/pJET) clone 4
pPS16_013 (FKBP/pJET) clone 5
pPS16_010 (FRB) clone 2
pPS16_010 (FRB) clone 3
pPS16_010 (FRB) clone 5
pPS16_002 (1.2) clone 1
pPS16_002 (1.2) clone 11
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.
