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HD®
F200 (200μL) and F500 (500μL)

FuGENEOVERVIEW

FuGENE® HD Transfection Reagent is a multi-component reagent that forms a complex
with DNA, then transports the complex into animal or insect cells. This reagent issuitable for use in media with or without serum, and for transient or stable transfection.
With FuGENE HD, there is no need to change media after an incubation with thetransfection mix because of its minimal cytotoxicity.

STORAGE & HANDLING• Store FuGENE HD at +4C with the lid tightly closed in its original glass vial. Ifthe reagent is stored below 0C temporarily, briefly warm it to 37C to dissolve anyprecipitate.
• Always bring FuGENE HD to room temperature and mix well prior to use.
• When pipetting FuGENE HD from its glass vial into any working reagent mixes, DO
NOT allow the reagent to contact the plastic walls of the tube. Do not use siliconizedpipette tips or tubes.

OPTIMIZING TRANSFECTION CONDITIONS• Use the LightSwitch™ Transfection Optimization Kit (Cat# TFXOPT).
• Use of low-passage cell lines is strongly recommended as highly-passaged cells
generally yield inconsistent transfections.
• Test a variety of uL FuGENE HD to ug DNA ratios ranging from 2:1 to 6:1.
• Other parameters to vary include the amount of plasmid DNA used, seeded celldensity, and duration of transfection.

RELATED PRODUCTS
Item

Catalog no.

LightSwitch™ Transfection Optimization Kit

TFXOPT

GoClones™ Promoters

S7- - - - -GoClones™ 3´UTR Controls

S89- - - -LightSwitch™ Assay Reagent™ (100 assays)

LightSwitch™ Assay Reagent™ (1,000 assays)
GoClones™ Promoter Controls
GoClones™ 3´UTRsLS010

LS100

S79- - - -S8- - - - -Genome In. Function Out.

Tech support: 877-222-9543

GENERAL TRANSFECTION WORKFLOW
NOTE: Details related to optimal seeding density, duration of transfection, and amountsof plasmid or FuGENE HD to add will vary widely by cell line, conditions, and assay
type. For specific GoClone transfection protocol examples, see our online resources pageat http://switchgeargenomics.com/resources/.
DAY 1: Seed cells so they will be 80%-90% confluent at the time of transfection.
For 96-well format, seed cells in 100uL total volume per well.

DAY 2:1) Pre-warm serum free media (like Invitrogen’s Opti-MEM) to 37C and bring FuGENE
HD to room temperature (mix well).
2) Calculate the amount of DNA, FuGENE HD, and Opti-MEM that will be added toeach transfection well. For 96-well format, the total volume added to each wellshould be 5μL. We suggest starting with 50–100ng of DNA per well and testing 3:1
and 6:1 ratios of FuGENE HD to DNA (μL:ug). To prepare transfection complexes
for other well formats, proportionally change the quantity of all components basedon the relative difference in surface area of the well size being used.
3) Mix plasmid DNA and Opti-MEM.
4) Add FuGENE HD to DNA + Opti-MEM mix without touching the sides of the tube.
Mix well.
5) Incubate the transfection complex for 30 minutes at room temperature.
6) Add the transfection complex to the cells by gently dripping the transfection mix
into the well.
7) Incubate the cells for 18–72 hours before assessing the results. Transfection times
will vary by cell type and assay type.
Quality Control
Function is assessed using a reporter gene assay in COS-7 cells. Activity must be at least
75% compared to a reference lot. Cell growth is assessed using a cell proliferation test
which is an indirect measure of cytotoxicity. Cells are also tested for overall viability. A
detailed Certificate of Analysis is available upon request.
Label License
You shall use this Product in accordance with all applicable laws, rules, and regulations.
You may not attempt to reverse engineer this Product. If you are not willing to accept
these conditions, the distributor from whom the Product was purchased will accept
the return of the unopened Product and provide you with a full refund. However, in theevent this Product is opened, you agree to be bound by the conditions of this limited usestatement.

Genome In. Function Out.