Hsa-miR-663b, its specific antagonist or an irrelevant synthetic RNA (Qiagen, Milan, Italy) were transfected, in triplicate, in CD40-activated CLL cells (106 cells) at 50 nmol/l, using the AMAXA nucleofection system, the human B-cell kit and the U-015 program (Lonza, Cologne, Germany). After 18 h of culture, RNA was extracted for mRNA analysis.
U 015 program
Manufactured by Lonza
The U-015 program is a laboratory equipment product offered by Lonza. It is designed to perform specific functions within a laboratory setting. The core function of the U-015 program is to [FUNCTION DESCRIPTION]. Further details about the intended use or interpretation of the product are not available.
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2 protocols using u 015 program
1
Hsa-miR-663b Antagonist Effects
2
Optimized Transfection Protocols for Various Cell Types
Check if the same lab product or an alternative is used in the 5 most similar protocols
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3.106 293T cells per condition were transfected with 10 μg of plasmid DNA into 10 cm Petri dishes using lipofectamine 2000 (Invitrogen) according to the protocol provided by the supplier. The cells were used 48 h later for further experiments.
5.105 HBMEC per cuvette were washed in PBS and resuspended in 100 μl of Cell Line Nucleofector Solutions V (Lonza). These cells were transfected by nucleofection using the U-015 program (Lonza) with 3–4 μg of DNA.
2.106 or 5.106 CEM cells per condition were washed in PBS and resuspended in 100 μl of Cell Line Nucleofector Solutions V (Lonza). These cells were transfected by nucleofection using the C-016 program (Lonza) with 5 or 10 μg of DNA, respectively.
10.106 human primary T cells per condition were washed in PBS and resuspended in 100 μl of Human T Cell Nucleofector Solution (Lonza). These cells were transfected by nucleofection using the U-014 program (Lonza) with 10 μg of DNA.
All cells transfected by nucleofection were used for functional assays after an overnight incubation time.
5.105 HBMEC per cuvette were washed in PBS and resuspended in 100 μl of Cell Line Nucleofector Solutions V (Lonza). These cells were transfected by nucleofection using the U-015 program (Lonza) with 3–4 μg of DNA.
2.106 or 5.106 CEM cells per condition were washed in PBS and resuspended in 100 μl of Cell Line Nucleofector Solutions V (Lonza). These cells were transfected by nucleofection using the C-016 program (Lonza) with 5 or 10 μg of DNA, respectively.
10.106 human primary T cells per condition were washed in PBS and resuspended in 100 μl of Human T Cell Nucleofector Solution (Lonza). These cells were transfected by nucleofection using the U-014 program (Lonza) with 10 μg of DNA.
All cells transfected by nucleofection were used for functional assays after an overnight incubation time.
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