Team:Heidelberg/Notebook/miMeasure

From 2010.igem.org

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(→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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===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
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5000 Hela and HEK293T 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 50ng.
Condition a: cotransfected with stuffer (salmon sperm DNA)  
Condition a: cotransfected with stuffer (salmon sperm DNA)  
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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.
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The cells were used for measurements on day three.
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===TECAN measurement===
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The
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Revision as of 19:01, 24 October 2010

miMeasure

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

5000 Hela and HEK293T 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 50ng.

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.

The cells were used for measurements on day three.


TECAN measurement

The

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