Team:Heidelberg/Notebook/miMeasure

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(Difference between revisions)
(Seeding and transfection of cells for TECAN, flow cytometry, plate reader and microscopy)
(Seeding and transfection of cells for TECAN, flow cytometry, plate reader and microscopy)
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5000 cells were seeded on day one in each well of the 96 well plate. Transfection of the constructs (M12-M22) with four different conditions were carried out on day two. The ratio of transfection is 1 (M construct) : 5 (stuffer/ miRsAg/ pcDNA5/ shRNA3) with a total amount   
5000 cells were seeded on day one in each well of the 96 well plate. Transfection of the constructs (M12-M22) with four different conditions were carried out on day two. The ratio of transfection is 1 (M construct) : 5 (stuffer/ miRsAg/ pcDNA5/ shRNA3) with a total amount   
 +
Condition a: cotransfected with stuffer (salmon sperm DNA)  
Condition a: cotransfected with stuffer (salmon sperm DNA)  
Condition b: costransfected with synthetic RNA miRsAg  
Condition b: costransfected with synthetic RNA miRsAg  
Condition c: cotransfected with empty pcDNA5
Condition c: cotransfected with empty pcDNA5
Condition d: cotransfected with synthetic shRNA3
Condition d: cotransfected with synthetic shRNA3
 +
A control consisting of the empty miMeasure plasmid (without binding site) was also cotransfected wit the same conditions a, b, c and d.
A control consisting of the empty miMeasure plasmid (without binding site) was also cotransfected wit the same conditions a, b, c and d.

Revision as of 18:58, 24 October 2010

miMeasure

Seeding and transfection of cells for TECAN, flow cytometry, plate reader and microscopy

5000 cells were seeded on day one in each well of the 96 well plate. Transfection of the constructs (M12-M22) with four different conditions were carried out on day two. The ratio of transfection is 1 (M construct) : 5 (stuffer/ miRsAg/ pcDNA5/ shRNA3) with a total amount

Condition a: cotransfected with stuffer (salmon sperm DNA) Condition b: costransfected with synthetic RNA miRsAg Condition c: cotransfected with empty pcDNA5 Condition d: cotransfected with synthetic shRNA3

A control consisting of the empty miMeasure plasmid (without binding site) was also cotransfected wit the same conditions a, b, c and d.