Transfection Reagents Product Brochure - Thermo Scientific

Thermo Scientific
Transfection Reagents
Delivery solutions
for a wide range of applications
• DNA and shRNA plasmid delivery
• siRNA and microRNA transfection
• Co-transfection of plasmid and small RNA
• Specialized delivery solutions for difficult-to-transfect cells

Transfection reagents
for every application
We offer a comprehensive portfolio of competitively priced,
proven transfection reagents for a wide range of applications.
Nucleic acid being
transfected:
DNA/shRNA
plasmid
siRNA or
microRNA
Co-transfection of
plasmid and small RNA
Plasmid
Recommended product:
Thermo
Scientific
TurboFect
Thermo
Scientific
DharmaFECT
Thermo Scientific
DharmaFECT Duo
Thermo Scientific
TurboFect in vivo
Transfectable cell types in vitro • • •
Appropriate for in vivo use
•
Works in the presence of serum • •
DharmaFECT transfection reagent 1 is the most all-purpose reagent we offer for siRNA and microRNA transfection. Consult
the DharmaFECT Cell Type Guide at thermoscientific.com/DharmaFECTCellTypeGuide to see if the DharmaFECT transfection
reagent 2, 3, or 4 is a recommended alternative for your cell type.
Specialized delivery solutions for
difficult-to-transfect cells.
Check out
the Selection
Guide Online
Visit:
thermoscientific.
com/transfection
Many cell types are not amenable to transfection reagents, resulting in poor delivery or
high cytotoxicity. We offer specialized products to enable DNA and siRNA delivery into
difficult-to-transfect cells.
Nucleic acid being
transfected:
Recommended product:
siRNA
Thermo Scientific
Accell siRNA
Viral vectors
(shRNA or ORF)
Thermo Scientific
Trans-Lentiviral Packaging Kits
Difficult-to-transfect cells in vitro • •
Appropriate for in vivo use • •
Best for use when:
Conventional siRNA
delivery is inefficient or
results in unacceptable
levels of cell death
Transduction of high-titer viral
particles for shRNA or ORF delivery
is desired for gene knockdown
or expression studies;
producing stable cell lines
2

Higher plasmid transfection
efficiency across cell lines
Thermo Scientific TurboFect Transfection Reagent
is the reagent of choice for shRNA and DNA plasmid
transfection applications.
•Transfection efficiency
•Mean fluorescence intensity
GFP Positive Cells (%)
HeLa Cells
100
75
50
25
0
NTC TurboFect Lipofect- FuGENE TM 6
amine
2000
2000
1500
1000
500
0
Mean Fluorescence Intensity
WEHI Cells
GFP Positive Cells (%)
50
40
30
20
10
0
NTC TurboFect Lipofect- FuGENE 6
amine
2000
Thermo Scientific TurboFect Transfection Reagent demonstrates high efficiency transfection across cell lines. Cells were transfected
with plasmid DNA encoding eGFP with the indicated transfection reagent (NTC; non-treated control). Transfections were performed
according to manufacturers’ recommendations and subsequent GFP expression was analyzed by flow cytometry.
1500
1000
500
0
Mean Fluorescence Intensity
GFP Positive Cells (%)
HepG2 Cells
50
40
30
20
10
0
NTC TurboFect Lipofect- FuGENE 6
amine
2000
1400
1200
1000
800
600
400
200
0
Mean Fluorescence Intensity
Highlights
• High transfection
efficiency to a wide
variety of cell types
• Minimal toxicity when
compared to other
transfection reagents
• Transfection can
be performed in
the presence or
absence of serum
• Simple three-step
transfection protocol
eGFP
eCFP
DsRed
Triple co-transfection of
HeLa cells with Thermo
Scientific TurboFect
Transfection Reagent.
HeLa cells were
co-transfected with a
1:1:1 mixture of plasmids
encoding for enhanced
green fluorescent
protein localized in the
cytoplasm, enhanced
cyan fluorescent protein
with nuclear localization
signal and monomeric
DsRed protein
with membrane
localization signal.
3

siRNA and microRNA transfection
Thermo Scientific DharmaFECT Transfection Reagents
are specifically formulated for delivery of small RNAs.
Experimental reproducibility is essential to RNAi studies, yet poor siRNA delivery or cellular
toxicity may confuse the interpretation of results. While DharmaFECT transfection reagent 1
is the most all-purpose formulation, recommended for 80% of cell lines tested, DharmaFECT
transfection reagent 2, 3, and 4 offer chemically distinct alternatives for optimization prior to
RNAi screening or for characterization of transfection in novel cell types.
Check the
DharmaFECT
Cell Type
Guide
Normalized mRNA Expression (%)
120
100
80
60
40
20
0
100 nM
25 nM
5 nM
1 nM
100 nM
25 nM
PPIB MAPK1 PPIB
MAPK1
DharmaFECT 1
5 nM
1 nM
100 nM
25 nM
5 nM
1 nM
100 nM
HiPerFect
25 nM
5 nM
1 nM
Visit:
thermoscientific.
com/DharmaFECT
CellTypeGuide
Highlights
• Ideal for single-gene
studies or high-throughput
siRNA screens
• Effective delivery for target
silencing at low siRNA
concentrations
• Low toxicity even at a
broad range of
experimental conditions
for maximum
experimental flexibility
• Four distinct formulations
to enable optimization
studies in novel cell types
• DharmaFECT target mRNA levels
• HiPerFect target mRNA levels
Cell viability
Thermo Scientific DharmaFECT Reagent provides highly efficient delivery at
low siRNA concentrations. HeLa cells were transfected with Thermo Scientific
SMARTpool reagents targeting Cyclophilin B (PPIB) or MAPK1 at concentrations of
1, 5, 25, and 100 nM using DharmaFECT 1 or HiPerFect Reagents. mRNA expression
(bars) was determined by branched DNA assay and cell viability (data points) was
determined by Thermo Scientific alamarBlue Reagent.
4

Multiple Thermo Scientific
DharmaFECT formulations ensure
siRNA transfection success.
No single transfection reagent effectively delivers siRNA to the
hundreds of different cell lines being used in RNAi experiments.
Why compromise delivery efficiency and cell viability with a
“one-size-fits-all” transfection reagent
A published third-party study 1 of 384-well siRNA transfection
optimization demonstrated DharmaFECT formulations
out performing four competitor lipid reagents in 9 of 10
cell lines when assessing overall cell viability and transfection
efficiency. Notably, each of the four DharmaFECT formulations
was found to be optimal in one or more cell line, demonstrating
the value of chemically unique formulations.
1. J. Borawski, A. Lindeman, Optimization Procedure for small interfering RNA
Transfection in a 384-well format. J. Biomoleculer Screening. 12, 819-834 (2007).
Cell Line
HCT116
HEK293
UMR
A549
Huh7
XMD5
HeLa
SKOV3
MDA-MB-231
Optimal Transfection Reagent
DharmaFECT 1
DharmaFECT 2
DharmaFECT 3
DharmaFECT 4
A673 Lipofectamine 2000
Published siRNA transfection optimization
results for 10 cell lines. Reagents tested included
DharmaFECT 1, 2, 3, 4, HiPerFect, TransIT-TKO,
Lipofectamine 2000, and Oligofectamine
Transfection Reagent.
Reliable co-transfection results
Thermo Scientific DharmaFECT Duo Transfection Reagent
for co-transfection of plasmid with microRNA or siRNA.
DharmaFECT Duo transfection reagent demonstrates successful co-transfection of plasmid along
with a microRNA mimic or siRNA targeting Rluc in a reporter system to assess microRNA function.
• Effective Rluc expression (Plasmid Alone and Plasmid + Control)
• Down-regulation of expressed Rluc by the microRNA mimic (Plasmid + miRNA mimic)
• Greater than 85% knockdown of Rluc expression by siRNA (Plasmid + siRNA)
150
Plasmid Alone
Plasmid + Control
Plasmid + miRNA Mimic
Plasmid + siRNA
Normalized RLuc Expression (%)
125
100
75
50
25
0
miR-28
reporter
miR-95
reporter
miR-30d
reporter
miR-105
reporter
Thermo Scientific DharmaFECT Duo Transfection Reagent
effectively delivers both plasmid and small RNA molecules.
The psiCHECK-2 vector (100 ng/well; Promega) with cloned
microRNA recognition sites for miR-28, miR-95, miR-30d
and miR-105, respectively, were complexed alone or with
RNAi reagents (10 nM) using DharmaFECT Duo transfection
reagent (0.2 uL/well) in MCF7 cells at 10,000 cells/well
(96-well plate). The RNAi reagents were Thermo Scientific
miRIDIAN Mimics (Negative Control, miR-28, miR-95,
miR-30d and miR-105 or siRNA (Renilla luciferase pool).
Renilla luciferase expression was assessed at 48 hours using
the Dual-Glo Luciferase Assay System (Promega) and
normalized to identically treated psiCHECK-2 empty vector.
5

Plasmid delivery to animal models
Thermo Scientific TurboFect in vivo Transfection
Reagent is highly effective for DNA plasmid delivery
in vivo, while not inducing an inflammatory response.
No prolonged inflammatory
response after DNA
delivery using Thermo
Scientific TurboFect
in vivo transfection
reagent. 50 µg of plasmid
DNA was delivered into
female BALB/c mice via
intravenous injection
using TurboFect in vivo
Transfection Reagent.
The concentration of
proinflammatory cytokine
TNFα was determined
by ELISA in blood serum
samples taken at the
indicated time points.
(LPS; lipopolysaccharides).
TNFα (pg/mL)
1,400
1,200
1,000
800
600
400
200
0
Negative control (DNA only)
Positive control (LPS)
TurboFect in vivo
Transfection Reagent
0 hours 1 hour 3 hours 6 hours 24 hours
Highlights
• No detectable
inflammatory response
• Successful plasmid
delivery via multiple
routes of administration
• Functionally tested in
mice and rats
Review
our online
Resource
Library
Visit:
thermoscientific
com/resourcelibrary
Specialized delivery solutions
Thermo Scientific Trans-Lentiviral Packaging kits
for generation of lentiviral particles.
Take advantage of the most advanced strategy for generating lentiviral particles for transduction of a gene or
shRNA of interest. Co-transfection of the Trans-Lentiviral packaging plasmid mix and transfer vector into the
packaging cell line, HEK293T, allows efficient production of lentiviral supernatant.
Packaging Cell
6
Packaging Mix
Transfer
Vector
Transfection
Viral Proteins
Transcription
RNA
Transduce Cells
Titer 1-5 x 10 6 TU/mL
Replicationincompetent
Virus
Assembly
& Budding
Harvest Viral
Supernatant
Highlights
• 1 to 5 ×10 6 transducing
units (TU) per mL are
commonly achieved in
collected supernatant
• Effectively transduce
most mammalian cell
lines, including primary
and non-dividing cells
• Contains all genetic
elements necessary to
package both second
and third generation
lentiviral vectors

Thermo Scientific Accell siRNA for delivery to
difficult-to-transfect cells without a transfection reagent.
Proprietary modifications to Accell siRNA enables effective delivery to cell types such as neurons,
immunological cells, beta-islet cells, and other primary cell types.
White
matter
Granular
layer
Purkinje
layer
Molecular
layer
Molecular layer
Purkinje layer
Granular layer
A.
B.
Thermo Scientific Accell siRNA demonstrates effective uptake and silencing in organotypic brain slices. Accell
siRNA shows increased uptake with extended incubation. 250 μM cerebellar sections were prepared, cultured
and incubated for A. 3 hours and B. 72 hours with Accell Red Non-targeting control (NTC) siRNA before inspection
by microscopy.
anti-
Cyclophilin B
(PPIB)
anti-Actin
Media only Non-targeting control Thermo Accell Scientific PPIB Accell PPIB
96 hours 48 48 hours 72 72 hours 96 96 hours 48 hours 72 72 hours 96 hours
anti-GAPDH
Non-targeting
control
Thermo Scientific
Accell GAPDH
72 hours 96 hours 48 hours 72 hours 92 hours
Highlights
• Pre-designed, genome-wide
siRNA reagents for human,
mouse, and rat
• Species-specific Control Kits
for assessment of Accell
technology in your cell type
• Convenient two-step delivery
protocol for faster results
Thermo Scientific Accell siRNA effectively silences target
genes in cultured P8 mouse brain slices. Cultured brain
slices were assessed following incubation with Accell siRNA
targeting Cyclophilin B (PPIB) and GAPDH or Non-targeting
control (NTC) at three time points (48, 72, 96 hours) and the
level of remaining protein was evaluated using western blot
analysis (Actin as loading control).
For more
Accell
information
Visit:
thermoscientific.
com/Accell
7

Order details
Product Quantity Cat #
TurboFect Transfection Reagent
1 mL R0531
5 × 1 mL R0532
0.2 mL T-2001-01
0.75 mL T-2001-02
DharmaFECT 1 siRNA Transfection Reagent
1.5 mL T-2001-03
5 × 1.5 mL tubes T-2001-04
2 × 10 mL vials T-2001-07A
0.2 mL T-2002-01
0.75 mL T-2002-02
DharmaFECT 2 siRNA Transfection Reagent
1.5 mL T-2002-03
5 × 1.5 mL tubes T-2002-04
2 × 10 mL vials T-2002-07A
0.2 mL T-2003-01
0.75 mL T-2003-02
DharmaFECT 3 siRNA Transfection Reagent
1.5 mL T-2003-03
5 × 1.5 mL tubes T-2003-04
2 × 10 mL vials T-2003-07A
0.2 mL T-2004-01
0.75 mL T-2004-02
DharmaFECT 4 siRNA Transfection Reagent
1.5 mL T-2004-03
5 × 1.5 mL tubes T-2004-04
2 × 10 mL vials T-2004-07A
0.2 mL T-2005-01
DharmaFECT Set of 4 siRNA Transfection Reagents
0.75 mL T-2005-02
1.5 mL T-2005-03
0.2 mL T-2010-01
DharmaFECT Duo Transfection Reagent
0.75 mL T-2010-02
1.5 mL T-2010-03
5 × 1.5 mL tubes T-2010-04
TurboFect in vivo Transfection Reagent 120 µL R0541
10 rxn kit TLP5912
Trans-Lentiviral shRNA Packaging Kit with Calcium Phosphate
50 rxn kit TLP5913
100 rxn kit TLP5914
Calcium Phosphate Transfection Reagent 50 rxn TLP5915
10 rxn kit TLP5916
Trans-Lentiviral ORF Packaging Kit with Calcium Phosphate
50 rxn kit TLP5919
100 rxn kit TLP5920
Trans-Lentiviral shRNA Packaging Kit with Calcium Phosphate and HEK293T 10 rxn kit TLP5917
Trans-Lentiviral ORF Packaging Kit with Calcium Phosphate and HEK293T 10 rxn kit TLP5918
• Learn more at
thermoscientific.com/
transfection
© 2013 Thermo Fisher Scientific Inc. All rights reserved. HiPerfect is a trademark of QIAGEN Inc. Lipofectamine and Oligofectamine are trademarks of Invitrogen
Corporation. TransIT-TKO is a trademark of Mirus Bio Corporation. FuGENE is a trademark of Fugent, L.L.C. psiCHECK and Dual-Glo are trademarks of Promega.
All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.
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Fax 00800 222 00 889
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Technical Support
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Tel 800 235 9880
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Technical Support
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