Team:Heidelberg/Notebook/miMeasure

From 2010.igem.org

(Difference between revisions)
(Seeding and transfection of cells for microscopy and flow cytometry)
Line 19: Line 19:
The cells were used for measurements on day three.
The cells were used for measurements on day three.
 +
[[Image:M12-M22_HeLa_daten.jpg]]

Revision as of 21:32, 26 October 2010

miMeasure

Seeding and transfection of cells for microscopy and flow cytometry

5000 Hela 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 of 50ng DNA.

Condition a: cotransfection with stuffer (salmon sperm DNA)

Condition b: cotransfection with synthetic RNA miRsAg

Condition c: cotransfection with empty pcDNA5

Condition d: cotransfection with synthetic shRNA3

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

The cells were used for measurements on day three. M12-M22 HeLa daten.jpg


July
MTWTFSS
1234
567891011
12131415161718
19202122232425
262728293031
August
MTWTFSS
1
2345678
9101112131415
16171819202122
23242526272829
3031
September
MTWTFSS
12345
6789101112
13141516171819
20212223242526
27282930 -
October
MTWTFSS
123
45678910
11121314151617
18192021222324
25262728293031