Silencer庐 Select Human Ion Channel siRNA Library V4 Product Information Sheet
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Product: Silencer Select Human Ion Channel siRNA Library V4
Catalog # (P/N): 4397915
Lot #: AMO01Z73
Amount: 0.25 nmol each siRNA
Content: 1014 unique siRNAs targeting each of 338 human ion channel genes*
Total of 12 96-well plates (plates are Axygen #PCR96FS; www.axygen.com)
9 plates with 88 siRNAs each
3 plates with 74 siRNAs each
*A few siRNAs have more than one gene target, due to gene families with highly homologous members or
predicted genes with high homology to verified genes. See the accompanying CD for annotation information
and siRNA details.
Purity: Standard
siRNA Format: Annealed
Appearance: Powder
Storage: Store at or below 鈥?20掳C. Do not store in a frost-free freezer. (Dried oligonucleotides are shipped at
ambient temperature.)
Shelf Life: 1 year, when stored at or below 鈥?20掳C.
USER INFORMATION
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Product Description: This Silencer Select siRNA Library is composed of Silencer Select Pre-designed and, where available,
Validated siRNAs, targeting human ion channel genes identified using the PANTHER鈩? and Gene Ontology
databases.
Silencer Select siRNAs are designed using a novel algorithm that was developed utilizing the latest in
machine-learning methods. These next-generation siRNAs exhibit up to 100-fold higher silencing potency
than siRNAs from other leading siRNA manufacturers. Off-target activity (assayed by microarray analysis) is
blocked by up to 90% because Silencer Select siRNAs can be used at 5- to 20-fold lower concentrations,
are bioinformatically screened using the latest knowledge about miRNA seed regions and toxic sequence
motifs, and incorporate strategic chemical modifications. As a result, Silencer Select siRNAs provide
unrivalled specificity and cleaner, more consistent phenotypic data.
Handling Instructions: RNA oligonucleotides are susceptible to degradation by exogenous ribonucleases introduced during
handling. Wear gloves when handling this product. Use RNase-free reagents, tubes, and barrier pipette tips.
Upon receipt, your siRNAs may be safely stored in a non-frost-free freezer at or below 鈥?20掳C (dried
oligonucleotides are shipped at ambient temperature).
Resuspension of siRNAs
Centrifuge each plate at low speed (maximum RCF 4,000 X g) to collect the contents at the bottom of the
wells before removing the seal.
Important: Perform this process in a tissue culture hood.
1. Remove seal carefully.
2. Add nuclease-free, sterile water, using a multichannel pipette and sterile tips, to achieve the desired
concentration.*
3. Gently pipet up and down 5 times to resuspend.
4. Centrifuge briefly to collect the liquid at the bottom of the wells, if necessary.
5. (Optional) Aliquot the siRNAs into one or more daughter plates, to limit the number of freeze-thaw
cycles to which the siRNAs are subjected.
6. Place a new sterile seal (such as Axygen PCR-AS-200) on the plate before storing.
7. Store at 鈥?80掳C until ready to use.
* An online calculator for suspension of dried oligonucleotides is available at
www.ambion.com/techlib/append/oligo_dilution.html
Ambion, Inc. 鈥? An Applied Biosystems Business 鈥? +1 800-888-8804 鈥? www.ambion.com 鈥? e-mail: techserv@ambion.com
Cat #4397915 Page 1 of 3
Lot # AMO01Z73 03/26/2008
Silencer庐 Select Human Ion Channel siRNA Library V4 Cat # 4397915
Applications: Transfecting Silencer Select siRNAs Into Mammalian Cells
The efficiency with which mammalian cells are transfected with siRNA will vary according to cell type and the
transfection agent used. This means that the optimal concentration used for transfections should be
determined empirically. Since Silencer Select siRNAs exhibit superior silencing potency compared to other
siRNAs, we suggest starting concentrations of 5- to 20-fold less than typically used for transfection of your
experimental cell lines. We have found that Silencer Select siRNAs reduced mRNA levels >80% at final
concentrations of 2鈥?10 nM using lipid-mediated transfection in HeLa and U-2 OS human osteosarcoma
cells.
a
General Transfection Starting Points for Mammalian Cells
Plate Format 384 wells 96 wells 24 wells 12 wells
b
0.3 碌L 0.3鈥?1.0 碌L 1鈥?3 碌L 2鈥?4 碌L
Transfection Agent
c
0.35鈥?0.5 pmol 0.5 pmol 2.5 pmol 5 pmol
siRNA
d
4000鈥?8000 cells/well 6000 cells/well 40,000 cells/well 80,000 cells/well
Cell Density
Final Volume per Well 70鈥?100 碌L 100 碌L 0.5 mL 1.0 mL
a
Appropriate for lipid-mediated transfection and easily transfected cell lines such as HeLa.
b
Refer to the instructions provided with your transfection agent for the recommended volume.
c
The siRNA amounts indicated result in a final siRNA concentration of 5 nM. The amount of siRNA required for maximal gene
silencing will vary among cell types. For a 96-well plate, and a 100 碌L final transfection volume, 0.5 pmol of a 1 碌M siRNA solution
is 0.5 碌L. Robotic pipettors may require volumes of 2鈥?5 碌L for accurate pipetting. To increase pipetting volumes and accuracy
when preparing transfection complexes, we recommend first preparing a plate with a dilution of your stock siRNA.
d
Optimal cell density will vary among cell types, depending on cell size and growth characteristics. In general, 30鈥?70% confluency
is recommended.
Transfection Optimization
Optimizing transfection efficiency is crucial for maximizing gene silencing while minimizing cytotoxicity.
Optimal transfection efficiencies are achieved by identifying an effective transfection agent for each cell type
and by adjusting (in order of importance):
鈥? Amount of transfection agent
鈥? Amount of siRNA
鈥? Cell density at the time of transfection
鈥? Order of transfection (pre-plating cells or plating cells/transfecting in tandem)
鈥? Length of exposure of cells to transfection agent/siRNA complexes
Most protocols recommend maintaining mammalian cells in the medium used for transfection; this avoids
diluting or removing the siRNAs from the cells by adding medium or washing the cells with new medium too
soon after transfection. We have found that cells typically exhibit greater viability when existing medium is
replaced by fresh medium 24 hours after transfection. Replacing medium after 24 hours generally does not
change the activity of the transfected siRNAs.
Once the conditions for maximal gene silencing are determined, they should be kept constant from
experiment to experiment for a given cell type. Include controls in all plates for each experiment to ensure
consistency.
For additional information about siRNA transfection, including transfection conditions for many cell types and
optimization protocols, see the siRNA Delivery Resource at:
www.ambion.com/techlib/resources/delivery
RELATED PRODUCTS
庐
Silencer Select Pre-designed and Validated siRNAs
Cat #Various (see www.ambion.com/geneassist)
An all-new class of modified siRNAs with unsurpassed efficacy, potency and specificity. Search the
GeneAssist鈩? Atlas at www.ambion.com/geneassist to find guaranteed-to-silence siRNAs to your gene of
interest.
庐
Silencer Select Control siRNAs
Cat #Various (see www.ambion.com/siRNA)
Validated, nontargeting siRNAs (Silencer Select Negative Controls) and proven-to-work positive control
siRNAs (Silencer Select GAPDH siRNAs) with the Silencer Select modifications.
siPORT鈩? NeoFX鈩? Transfection Agent
Cat #AM4510 and AM4511
A versatile lipid-based agent for efficient and reproducible transfection of adherent cells while subculturing,
without increased cytotoxicity.
庐
TaqMan Gene Expression Assays
See www.allgenes.com or www.ambion.com/geneassist
A comprehensive collection of over 700,000 probe and primer sets for quantitative gene expression analysis
using real-time PCR. Search the GeneAssist鈩? Atlas at www.ambion.com/geneassist to find suggested
TaqMan Gene Expression Assays for the gene targeted by an siRNA of interest.
Ambion, Inc. 鈥? An Applied Biosystems Business 鈥? +1 800-888-8804 鈥? www.ambion.com 鈥? e-mail: techserv@ambion.com
Cat #4397915 Page 2 of 3
Lot # AMO01Z73 03/26/2008
Silencer庐 Select Human Ion Channel siRNA Library V4 Cat # 4397915
QUALITY CONTROL
Identity: The mass of a sample of each single-stranded RNA oligonucleotide is analyzed using MALDI-TOF mass
spectrometry and compared to the calculated mass.
Annealing: A sample of the annealed siRNA is analyzed by nondenaturing gel electrophoresis.
OTHER INFORMATION
Material Safety Data Sheets (MSDSs) can be printed or downloaded from product-specific links on our website at the
Material Safety Data
following address: www.ambion.com/techlib/msds. Alternatively, e-mail your request to
Sheets:
MSDS_Inquiry_CCRM@appliedbiosystems.com. Specify the catalog or part number(s) of the product(s), and we will e-mail
the associated MSDSs unless you specify a preference for fax delivery. For customers without access to the internet or fax,
our technical service department can fulfill MSDS requests placed by telephone or postal mail. (Requests for postal delivery
require 1鈥?2 weeks for processing.)
For research use only. Not for use in diagnostic procedures.
Warranty and Liability:
Applied Biosystems is committed to delivering superior product quality and performance, supported by industry-leading global
service and technical support teams. Warranty information for the accompanying consumable product is available at
www.ambion.com/info/warranty in "Limited Warranty for Consumables," which is subject to the exclusions, conditions,
exceptions, and limitations set forth under the caption 鈥淓XCLUSIONS, CONDITIONS, EXCEPTIONS, AND LIMITATIONS鈥? in
the full warranty statement. Please contact Applied Biosystems if you have any questions about our warranties or would like
information about post-warranty support.
Information in this document is subject to change without notice. Applied Biosystems assumes no responsibil-ity for any errors
that may appear in this document.
Applied Biosystems disclaims all warranties with respect to this document, expressed or implied, including but not limited to
those of merchantability or fitness for a particular purpose. In no event shall Applied Biosystems be liable, whether in contract,
tort, warranty, or under any statute or on any other basis for special, incidental, indirect, punitive, multiple or consequential
damages in connection with or arising from this document, including but not limited to the use thereof.
Applied Biosystems, AB (Design), Ambion, and Silencer are registered trademarks and GeneAssist, NeoFX, PANTHER,
Trademarks, Patents
and siPORT are trademarks of Applera Corporation or its subsidiaries in the US and/or certain other countries. TaqMan is a
and Licensing:
registered trademark of Roche Molecular Systems, Inc. LNA is a registered trademark of Exiqon A/S, Vedbaek, Denmark.
All other trademarks are the sole property of their respective owners.
NOTICE TO PURCHASER:
This product is licensed under European Patents 1144623, 1214945 and foreign equivalents from Alnylam Pharmaceuticals,
Inc., Cambridge, USA and is provided only for use in academic and commercial research including in vitro and in vivo
identification and validation of drug targets (but excluding the evaluation or characterization of this product as the potential
basis for a siRNA-based drug) and not for any other commercial purposes. Information about licenses for commercial use
(including discovery and development of siRNA-based drugs) is available from Alnylam Pharmaceuticals, Inc., 300 Third
Street, Cambridge, MA 02142, USA.
Ambion siRNA products are manufactured under license from the Massachusetts Institute of Technology to U.S. Patent
Nos. 7,056,704 and 7,078,196 and pending counterparts.
Purchase of this product gives the user rights under U.S. Patent No. 6,506,559, and its foreign equivalents, to use this
product for life science research, not for use in humans or clinical diagnostics.
Locked Nucleic Acids are covered by patents and patent applications owned by Exiqon A/S. This product is for research use
only and not for diagnostic or therapeutic use. The products may be used only for the buyer's internal research purposes
and not for commercial use except for providing a service to research customers. The buyer may not resell products in their
original or any modified form.
A license to perform the patented 5鈥? Nuclease Process for research is obtained by the purchase of (i) both Licensed Probe
and Authorized 5' Nuclease Core Kit, (ii) a Licensed 5鈥? Nuclease Kit, or (iii) license rights from Applied Biosystems.
The TaqMan庐 Gene Expression Assays contain Licensed Probe. Use of the TaqMan Gene Expression Assays is covered by
one or more of the following US patents and corresponding patent claims outside the US: 5,538,848, 5,723,591, 5,876,930,
6,030,787, 6,258,569, and 5,804,375 (claims 1-12 only). The purchase of this product includes a limited, non-transferable
immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser鈥檚 own internal
research. The right to use this product in the 5鈥? Nuclease Process under the applicable claims of US Patents Nos. 5,210,015
and 5,487,972, and corresponding patent claims outside the United States, can be obtained through purchase of an
Authorized 5' Nuclease Core Kit. Except under separate license rights available from Applied Biosystems, no right under any
other patent claim, or to perform commercial services of any kind, including without limitation reporting the results of
purchaser's activities for a fee or other commercial consideration, or to sublicense, repackage with other products, or resell in
any form, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Human in vitro
diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may
be obtained from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
漏 2008 Ambion, Inc. All rights reserved. 4398632A
Ambion, Inc. 鈥? An Applied Biosystems Business 鈥? +1 800-888-8804 鈥? www.ambion.com 鈥? e-mail: techserv@ambion.com
Cat #4397915 Page 3 of 3
Lot # AMO01Z73 03/26/2008
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