Magnetofection™

PolyMag & PolyMag Neo

INSTRUCTION MANUAL

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Magnetofection: PolyMag, PolyMag Neo

Instruction Manual

Magnetofection is a simple and highly efficient in vitro and in vivo transfection method.

List of Magnetofection™ Kits

Catalog Description Volume (µL) Size (number of Number of

Number transfections / µg of transfections /
DNA) 96 well plates
PN30100 PolyMag reagent 100 100 1000
PN30200 PolyMag reagent 200 200 2000
PN31000 PolyMag reagent 1000 1000 10000
PG60100 PolyMag neo reagent 100 100 1000
PG60200 PolyMag neo reagent 200 200 2000
PG61000 PolyMag neo reagent 1000 1000 10000
KC30200 Magnetofection Starting Kit 1 3 x 100 200 2000
KC30300 SiRNA Starting kit 2 200 +2x100
KC30400 Super Starting Kit 3 200 + 3 x 100 200 400 - 2000
MF10000 Super Magnetic Plate N/A N/A N/A
MF14000 Mega Magnetic Plate N/A N/A N/A
MF10096 96-Magnets, Magnetic Plate N/A N/A N/A
1
Contains 1 vial of each reagent (PolyMag, PolyMag Neo and CombiMag) + one Super Magnetic Plate (MF10000)
2
Contains 1 vial of each reagent (SilenceMag, PolyMag PolyMag Neo) + one Super Magnetic Plate (MF10000)
3
Contains 1 vial of each reagent (PolyMag, PolyMag Neo, CombiMag and SilenceMag) + one Super Magnetic Plate (MF10000)

Use the content of the table above to determine the appropriate catalog number for your needs. You can order
these products by contacting us (phone, fax, email, website). Several kits containing different Magnetofection
reagents are also available, please contact us (contact@ozbiosciences.com) for a more detailed offer. For all other
supplementary information, do not hesitate to contact our dedicated technical support (tech@ozbiosciences.com)
and/or to visit our website: www.ozbiosciences.com.

OZ Biosciences SAS OZ Biosciences INC

163 avenue de Luminy 4901 Morena Blvd,
Case 922, zone entreprise Suite 501
13288 Marseille cedex 09 - FRANCE San Diego CA 92117 - USA
Ph: +33 (0) 486 948 516 Ph : + 1-858-246-7840
Fax: +33 (0) 486 948 515 Fax : + 1-855-631-0626
contact@ozbiosciences.com contactUSA@ozbiosciences.com
order@ozbiosciences.com orderUSA@ozbiosciences.com

www.ozbiosciences.com

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Table of Contents

1. Technology 2-3
1.1. Description 2
1.2. Available Reagents 2
1.3. Kit Contents, Stability and Storage 3
2. Applications 3
3. Magnetofection Apparatus 3
4. Protocols 4-5
4.1. General Considerations 4
4.2. Cell preparation 4
4.3. Transfection protocol 4
4.4. Magnetofection of Suspension Cells 5
5. Appendix 6
5.1. Protocol Optimization 6
5.2. Quality Controls 6
5.3. Troubleshooting 6
6. Related Products 7
7. Purchaser Notification 8

1. Technology

1.1. Description

Magnetofection™ is an original, simple and highly efficient method to transfect cells in vitro and in vivo. It
exploits magnetic force exerted upon gene vectors associated with magnetic particles to drive the vectors
towards, possibly even into, the target cells. In this manner, the complete applied vector dose gets concentrated
onto cells within a few minutes so that 100% of the cells get in contact with a significant vector dose.

This has several important consequences:

1. Greatly improved transfection rates in terms of percentage of transfected cells compared to standard
transfection.
2. Up to several thousand folds increased levels of transgene expression compared to standard transfection.
3. High transfection rates and transgene expression levels are achievable with extremely low vector doses,
which allow saving expensive transfection reagents.
4. Extremely short process time in comparison to standard procedures. A few minutes of incubation of cells
with gene vectors are sufficient to generate high transfection efficiency.

Based upon a validated and recognized magnetic drug targeting technology, this innovative method is:
• Efficient, simple & rapid
• Multipurpose (for all types of nucleic acids and non-viral vectors)
• Universal (primary cells and cell lines)
• Non toxic & economical

1.2. Available Reagents

OZ Biosciences offers different ready-to-use Magnetofection transfection reagents for in vitro applications:
1. PolyMag is a universally applicable magnetic particle preparation for high efficiency nucleic acid delivery.
Nucleic acids to be transfected and the magnetic particles are mixed in a one-step procedure. PolyMag has
been used successfully with plasmid DNA, antisense oligonucleotides and siRNA.
2. PolyMag Neo is an optimized formulation of PolyMag for a higher gene expression level in primary, hard-to-
transfect and cell lines.

PolyMag is also available in vivo grade (in vivo PolyMag) for your targeted gene delivery in vivo. Further detailed
information on Magnetofection™ reagents can be found at: www.ozbiosciences.com

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 1.3. Kit Contents, Stabiblity and Storage

Kit contents differ according to their size

1 tube containing 100 µL of particle suspension good for 100 transfections with 1 µg of DNA
1 tube containing 200 µL of particle suspension good for 200 transfections with 1 µg of DNA
1 tube containing 1000 µL of particle suspension good for 1000 transfections with 1 µg of DNA

Stability and Storage

Storage: +4ºC. Upon receipt and for long-term use, store all reagent tubes in the fridge. Magnetofection reagents
are stable for at least one year at the recommended storage temperature.

• DO NOT FREEZE THE MAGNETIC NANOPARTICLES!

• DO NOT ADD ANYTHING TO THE STOCK SOLUTION OF NANOPARTICLES!

Shipping condition: Room temperature

2. Applications

Magnetofection is applicable with numerous cell types and has been successfully tested on a variety of
immortalized cell lines as well as primary cells (see list on website). If a particular cell type or cell line is not
listed, this does not imply that Magnetofection is not going to work. An updated list of cells successfully
tested as well as product citations is available on the website: www.ozbiosciences.com.

3. Magnetofection Apparatus

Besides suitable magnetic nanoparticles, Magnetofection requires an appropriate magnetic field generated by
a magnetic plate especially designed for Magnetofection. Its special geometry not only produces strong
magnetic fields under each well of 96-well plates but is also applicable to other plate formats (T-75 flasks, 60 &
100 mm dishes, 6-, 12- and 24-well plates). Super Magnetic Plate suits for all cell culture supports and Mega
Magnetic Plate is designed to hold up to 4 culture dishes at one time.

Magnetic plate 96 magnets Super Magnetic Plate Mega Magnetic Plate

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4. Protocols

4.1. General Considerations

Instructions given below represent sample protocols that were successfully applied to a variety of cell lines.
Optimal conditions do vary from cell line to cell line and are dependent on nucleic acid used. Consequently, the
amounts and ratio of the individual components (DNA and reagent) may have to be adjusted to achieve best
results. Therefore, we advise you to optimize the various transfection parameters (components concentration,
cell number, incubation time…). Several protocol optimizations are available in the Appendix and upon request
by email. The following recommendations can be used as guidelines as starting point to achieve good
transfection.

4.2 Cells Preparation

It is recommended to seed or plate the cells the day prior transfection. The suitable cell density will depend on
the growth rate and the cells conditions. Cells should be 60-90% confluent at the time of Magnetofection (see
the suggested cell number in the table below). For suspension cells, use the specific protocol given below.
Immediately preceding transfection, the medium can be replaced with fresh medium (optionally without
serum).
Table 1: Cell Number and Transfection Volume Suggested
Tissue Culture Dish Cell Number DNA Quantity Transfection
(µg) Volume
96 well 0.5 – 2 x 10 4 0.1 – 0.5 200 µL
24 well 0.5 – 1 x 10 5 0.5 - 2 500 µL
12 well 1 – 2 x 10 5 2-4 1 mL
6 well 2 – 4 x 10 5 2-6 2 mL
60 mm dish 5 – 10 x 10 5 6-8 4 mL
90 - 100 mm dish 10 – 20 x 10 5 8 - 12 8 mL
T-75 flask 20 – 50 x 10 5 10 - 20 12 mL

4.3. Transfection protocol

Use the following procedure to transfect DNA into mammalian cells. The Table 1 shows transfection condition
according to different cell culture formats (all amount are given on per-well basis). The DNA and the
magnetofection reagent (PolyMag or PolyMag Neo) should be at room temperature and be gently vortexed
prior to use. The protocol is as simple as follows: use 1 µL of PolyMag or PolyMag Neo per µg of DNA.

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1) Before each use, vortex the PolyMag or PolyMag Neo tube. Add 1 to 10 µL of PolyMag or PolyMag Neo
(according to the DNA amount) to a microtube or to a microwell (U-bottom well is preferred to get a better
mixing). If required and for doses less than 1 µL, predilute PolyMag or PolyMag Neo with deionized water.

2) Dilute 1 to 10 µg of DNA to 200 µL with serum and supplement-free culture medium (such as DMEM).

3) Add the 200 µL DNA solution to the PolyMag or PolyMag Neo solution and mix immediately by vigorous
pipetting.

4) After 20 to 30 minutes of incubation, add the transfection mix (DNA + PolyMag or PolyMag Neo) to the
cells. The total transfection volume per well (culture medium + PolyMag or PolyMag Neo mixture) is
suggested in the Table 1.

5) Place the cell culture plate upon the magnetic plate for 5 to 20 minutes.

6) Optionally perform a medium change and then remove the magnetic plate.

7) Cultivate the cells under standard conditions until evaluation of transgene expression

The same protocol can be used to produce stably transfected cells except that 48 hours post-transfection fresh
medium containing the appropriate antibiotics are transferred to cells for selection. It is important to wait at
least 48 hours before exposing the transfected cells to selection media.

For most cell types, a medium change is not required after Magnetofection. However, it may be necessary for
cells that are sensitive to serum/supplement concentration. Alternatively, the cells may be kept in serum-free
medium during Magnetofection (up to 4 hours). In this case, a medium change will be required after
Magnetofection.

4.4. Magnetofection of suspension cells

1) The composition and preparation of PolyMag or PolyMag Neo / DNA are performed exactly as described
above from steps 1 to 3.

2) While PolyMag or PolyMag Neo / DNA are incubating (step 4 above), dilute the cells to be transfected to 5
x 105 - 1 x 106 / mL in medium (with or without serum- or supplement; depending on cell type and
sensitivity of cells towards serum-free conditions) and perform one of the following three options to
sediment the cells at the bottom of the culture dish in order to promote the contact with the magnetic
nanoparticles.

a. Seed the cells on polyLysine-coated plates and use the protocol for adherent cells.
OR
b. Briefly, centrifuge the cells (2 minutes) to pellet them and use the protocol for adherent cells.
OR
c. Mix cell suspension with 30 µL of CombiMag (from OZ Biosciences) reagent per mL of cell suspension.
i. Incubate for 10 - 15 minutes.
ii. Distribute cells to your culture dish placed upon the magnetic plate.
iii. Incubate for 15 minutes
OR
d. Incubate the cells in serum free medium during 2 hours prior Magnetofection. The absence of serum
allows some cells to adhere onto the plastic dish surface.

3) Add the resulting mixture of PolyMag or PolyMag Neo / DNA to the cells while keeping the cell culture
plate on the magnetic plate.

4) Incubate for 15 minutes.

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5) Carefully remove the medium supernatant from the cells and replace with fresh complete medium while
the culture plate remains positioned on the magnetic plate. Be careful not to aspirate the magnetically
sedimented cells.

6) Remove culture plate from magnetic plate.

7) Continue to cultivate cells as desired until evaluation of transgene expression.

5. Appendix

5.1. Protocol Optimization

We strongly advise you to optimize your transfection conditions in order to get the best out of
Magnetofection. Several parameters can be optimized:
• Nucleic acid dose used
• Ratio of PolyMag or PolyMag Neo to nucleic acid
• Cell density
• Incubation time

OZ Biosciences team has investigated numerous factors during the course of Magnetofection reagent
development. Based on our experience, we recommend that you optimize one parameter at a time and start
from the experimental procedure described above in section 4.

1) Start by optimizing the ratio PolyMag or PolyMag Neo / DNA . To this end, use a fixed amount of DNA. Vary
the amount of PolyMag or PolyMag Neo from 0.25 to 5µL / µg of DNA. The ratio PolyMag or PolyMag Neo
/ DNA can be changed by doubling or multiplying the volume of the reagent used. Reagent can be pre-
diluted in deionized water.

2) Thereafter, change the nucleic acid dose with a fixed ratio of PolyMag or PolyMag Neo / DNA that has been
previously optimized. For this purpose, you can perform a serial dilution of a preformed magnetic vector
complex.

3) After having identified the correct quantities of PolyMag or PolyMag Neo and nucleic acid, you can pursue
the process by optimizing the cell number as well as the incubation times for the complex formation and for
the magnetic field application.

5.2. Quality Controls

To insure the performance of each lot of Magnetofection produced, we qualify each component using rigorous
standards. The following assays are conducted in vitro to qualify the function, quality and activity of each kit
component.
Components Standard Quality Controls
PolyMag, 1. Quality and size homogeneity of the magnetic nanoparticles.
PolyMag Neo 2. Stability of the magnetic nanoparticle formulations.
3. Transfection efficacies on NIH-3T3 and COS 7 cells. Every lot shall have an acceptance
specification of > 80% of the activity of a reference lot
Magnetic Plate 1. Tests of solidity
2. Test of the magnetic field force

5.3. Troubleshooting

Our dedicated and specialized technical support group will be pleased to answer any of your requests and to
help you with your transfection experiments. tech@ozbiosciences.com
An updated list of products citations is available on the website: www.ozbiosciences.com

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6. Related Products

Description
MAGNETOFECTION TECHNOLOGY
Super Magnetic Plate (standard size for all cell culture support)
Mega Magnetic plate (mega size to hold 4 culture dishes at one time)
Transfection reagents:
PolyMag Neo (for all nucleic acids)
Magnetofectamine™ kit: Lipofectamine™ 2000 + CombiMag (for all nucleic acids)
NeuroMag (dedicated for neurons)
SilenceMag (for siRNA application)
Transfection enhancer:
CombiMag (to improve any transfection reagent efficiency)
Viral Transduction enhancers:
ViroMag (to optimize viral transduction)
ViroMag R/L (specific for Retrovirus and Lentivirus)
AdenoMag (for Adenoviruses)
In vivo Magnetofection
In vivo ViroMag (for magnetic assisted viral infection)
In vivo PolyMag (polymer-based magnetic nanoparticles)
In vivo DogtorMag (lipid-based magnetic nanoparticles)
In vivo SilenceMag (for siRNA application)
LIPOFECTION TECHNOLOGY (LIPID-BASED)
Lullaby (siRNA transfection reagent)
DreamFect Gold (Transfection reagent for all types of nucleic acids)
VeroFect (for Vero cells)
Ecotransfect (Economical reagent for routine transfection)
FlyFectin (for Insect cells)
i-MICST TECHNOLOGY
Viro-MICST (to transduce directly on magnetic cell purification columns)
3D TRANSFECTION TECHNOLOGY
3DfectIN (for hydrogels culture)
3Dfect (for scaffolds culture)
RECOMBINANT PROTEIN PRODUCTION
HYPE-5 Transfection Kit (for High Yield Protein Expression)
PROTEIN DELIVERY SYSTEMS
Ab-DeliverIN (delivery reagent for antibodies)
Pro-DeliverIN (delivery reagent for protein in vivo and in vitro)
PLASMIDS PVECTOZ
pVectOZ-LacZ / pVectOZ-SEAP / pVectOZ-GFP / pVectOZ-Luciferase
ASSAY KITS
Bradford – Protein Assay Kit
MTT cell proliferation kit
β-Galactosidase assay kits (CPRG/ONPG)
BIOCHEMICALS
D-Luciferin, K+ and Na+ 1g
G-418, Sulfate 1g
X-Gal powder 1g

Please, feel free to contact us for all complementary information and remember to visit our website
(www.ozbiosciences.com) to stay informed on the latest breakthrough technologies and updated on our
complete product list.

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Purchaser Notification

Limited License
The purchase of the Magnetofection™ Reagent grants the purchaser a non-transferable, non-exclusive
license to use the kit and/or its separate and included components (as listed in section 1, Kit Contents). This
reagent is intended for in-house research only by the buyer. Such use is limited to the transfection of
nucleic acids as described in the product manual. In addition, research only use means that this kit and all of
its contents are excluded, without limitation, from resale, repackaging, or use for the making or selling of
any commercial product or service without the written approval of OZ Biosciences.

Separate licenses are available from OZ Biosciences for the express purpose of non-research use or
applications of the Magnetofection™ Reagent. To inquire about such licenses, or to obtain authorization to
transfer or use the enclosed material, contact the Director of Business Development at OZ Biosciences.

Buyers may end this License at any time by returning all Magnetofection™ Reagent material and
documentation to OZ Biosciences, or by destroying all Magnetofection™ Reagent components. Purchasers
are advised to contact OZ Biosciences with the notification that a Magnetofection™ Reagent kit is being
returned in order to be reimbursed and/or to definitely terminate a license for internal research use only
granted through the purchase of the kit(s).

This document covers entirely the terms of the Magnetofection™ Reagent research only license, and does
not grant any other express or implied license. The laws of the French Government shall govern the
interpretation and enforcement of the terms of this License.

Product Use Limitations

The Magnetofection™ Reagent and all of its components are developed, designed, intended, and sold for
research use only. They are not to be used for human diagnostic or included/used in any drug intended for
human use. All care and attention should be exercised in the use of the kit components by following proper
research laboratory practices.
For more information, or for any comments on the terms and conditions of this License, please contact:

Director of Business Development

OZ Biosciences
Parc Scientifique et Technologique de Luminy
Zone Luminy Entreprise, case 922
13288 Marseille Cedex 9, France
Ph: +33 (0)4.86.94.85.16
Fax: +33 (0)4.86.94.85.15
E-mail: business@ozbiosciences.com

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 CONTACTS

OZ Biosciences SAS OZ Biosciences INC

163 avenue de Luminy 4901 Morena Blvd,
Case 922, zone entreprise Suite 501
13288 Marseille cedex 09 San Diego CA 92117
FRANCE USA

Ph: +33 (0) 486 948 516 Ph : + 1-858-246-7840

Fax: +33 (0) 486 948 515 Fax : + 1-855-631-0626

contact@ozbiosciences.com contactUSA@ozbiosciences.com
order@ozbiosciences.com orderUSA@ozbiosciences.com
tech@ozbiosciences.com techUSA@ozbiosciences.com

www.ozbiosciences.com

Follow us!

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