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Neon transfection system cell protocols For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. © 2009 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses . W-085888 0609 www.invitrogen.com PBMC This protocol requires Buffer T for Primary Cells. Cell information Cell type/Description PBMC (peripheral blood mononuclear cell)/Primary cell Characteristics/Species Suspension/Human Tissue origin Blood Media RPMI 1640 with L-glutamine and 10% FBS Morphology Round single cell Doubling time NA Subculturing Medium renewal every 2 to 3 days Culture conditions Temperature: 37°C; atmosphere: air 95%/carbon dioxide (CO 2 ) 5% Electroporation parameters Pulse voltage (v) Pulse width (ms) Pulse number Cell density (cells/mL) Transfection Viability Tip type 2,150 20 1 2 x 10 7 23% 95% 10 μL Peripheral blood mononuclear cells (PBMCs) were transfected using the Neon™ transfection system and 1.0 μg of a plasmid encoding EGFP. 24 hours posttransfection, the cells were analyzed by light (A) and fluorescence microscopy (B). A B

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Neon™transfection system

cell protocols

For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. © 2009 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. W-085888 0609

www.invitrogen.com

PBMCThis protocol requires Bu�er T for Primary Cells.

Cell informationCell type/Description PBMC (peripheral blood mononuclear cell)/Primary cell

Characteristics/Species Suspension/Human

Tissue origin Blood

Media RPMI 1640 with L-glutamine and 10% FBS

Morphology Round single cell

Doubling time NA

Subculturing Medium renewal every 2 to 3 days

Culture conditions Temperature: 37°C; atmosphere: air 95%/carbon dioxide (CO2) 5%

Electroporation parametersPulse

voltage (v)Pulse

width (ms)Pulse

numberCell density

(cells/mL)Transfection

ViabilityTip

type

2,150 20 1 2 x 107 23% 95% 10 µL

Peripheral blood mononuclear cells (PBMCs) were transfected using the Neon™ transfection system and 1.0 µg of a plasmid encoding EGFP. 24 hours posttransfection, the cells were analyzed by light (A) and fluorescence microscopy (B).

A B