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amaxa eNews #1
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Nucleofector 96-well Shuttle System
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Nucleofection enters the High Throughput Arena
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The 96-well Shuttle is an add-on to the Nucleofector II Device* and is thus based on the same unique features as the existing single-cuvette based Nucleofector technology. By applying its unmatched Nucleofector technology to the new area of high throughput transfection, amaxa has again combined superior technology with simplicity and ease-of-use.
* Nucleofector I users can benefit from our special trade-in offer to upgrade to the Nucleofector II.
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The Nucleofector 96-well Shuttle System at a Glance
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The Nucleofector 96-well Shuttle System comprises the Nucleofector II Device (left side, device in the back), which serves as the program delivery unit, the 96-well Shuttle (left side, device in the front) and a laptop computer. Via the 96-well Shuttle Software the laptop contols the interaction between Nucleofector II Device and the 96-well Shuttle. The system works in conjunction with Nucleofector Kits and Protocols specifically designed for the 96-well Shuttle.
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The Nucleofector 96-well Shuttle System comes with newly developed disposable and modular Nucleocuvette plates, completely intuitive software, a small footprint and full liquid handling compatibility designed to excel in high throughput applications, such as siRNA-based target validation. Other applications in focus are the transfection of cDNA libraries, the generation of stable clones and protein production in difficult-to-transfect suspension cell lines. No matter which application you have in mind, amaxa provides you not only with the technology but also with profound and comprehensive information as well as support around the transfection process. We want to help you safely guide your experiment from cell culture to analysis of your transfection results.
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Your Experiments will Benefit from
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| High efficiencies | Up to 95% transfection efficiency with DNA combined with viabilities >90% | | | Up to 99% siRNA duplex transfer even in suspension cells
| | High reproducibility | Low intra- and inter-plate variance | | | No edge effects and no lot-to-lot variances
| | Access to difficult-to-transfect cell types | Neurons, T cells and other difficult-to-transfect cells types can be transfected
| | Ease of use | Optimized 96-well Nucleofector Kits and protocols for many primary cells and difficult-to-transfect cell lines
| | Substrate flexibility | Identical transfection conditions for DNA, siRNA, shRNA...
| | Transfection flexibility | Modular 2 x 8 Nucleocuvette plate for scalable throughput | | | Up to 96 independent programs applicable per plate | | | Variable cell numbers - from 104 to 106 cells per reaction
| | Rapid optimization | Optimization of any difficult-to-transfect cell line in just one plate
| | Safety | Disposable sterile Nucleocuvette plates minimizes the risk of cross contamination | | | Innovative conductive polymer - no metal ion release
| | HTS-compatibility | Automatic processing of a 96-well plate in 3-4 min | | | Meets prerequisites for integration into Liquid Handling Systems
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High efficiencies – Even with difficult-to-transfect cell types
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Several applications, such as target identification and validation, clearly imply the demand for high throughput transfection. With lipofection, efficient transfection is mainly restricted to easy-to-transfect cell lines, e.g. CHO or COS. With the power of Nucleofection it is possible to efficiently transfect difficult-to-transfect cell lines and non-dividing primary cells, such as resting T Cells or Neurons. As a result, target identification and validation can now be performed in virtually any cell, ensuring much more relevant results as the most appropriate biological test sytem can be chosen.
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Figure 1: Example for the transfection of primary cells using the 96-well Shuttle. Cells were transfected with 400 ng pmaxGFP and analyzed for maxGFP expression by flow cytometry 24h post nucleofection. Cell viabilities (PI staining) ranged between 60% and 92% depending on the cell type.
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Figure 2: Comparable transfection efficiencies using the 96-well Shuttle and the Nucleofector II. Cell lines (ATCC) were transfected with 400 ng pmaxGFP (2 µg for Nucleofector II) and analyzed for maxGFP expression by flow cytometry 24h post nucleofection. Cell viabilities (PI staining) ranged between 60% and 97%, depending on the cell line.
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Flexibility combined with safety and convenience: The 96-well Nucleocuvette
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96-well nucleofection occurs in the patent pending Nucleocuvette modules with electrodes consisting of an innovative conductive polymer material. Hence, no metal ions are released into the cell suspension during nucleofection. Six modules are combined in a frame to form a complete 96-well Nucleocuvette plate with SBS standard dimensions that can be handled by any robotic system. The modules are disposable - minimizing the risk of cross contamination – and can be used individually, allowing for scalable throughput. Each well can be addressed separately during the nucleofection process which allows for a complete optimization procedure testing up to 96 different conditions within one plate. Finally the transfection volume per well is only 20 µl which drastically reduces the number of cells (as low as 2 x 104, depending on the cell type) needed per experiment.
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Figure 3: 96-well Nucleocuvette modules and plates.96-well nucleofection occurs in disposable 2x8 Nucleocuvette modules which consist of an innovative conductive polymer material. Six modules are combined to form a complete 96-well Nucleocuvette plate
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Total Substrate Flexibility: One technology for siRNA, cDNA, and shRNA
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Just as the existing Nucleofector technology, the 96-well Shuttle system offers high flexibility with respect to applications, since identical transfection parameters apply for any nucleic acid substrate used. DNA vectors (e.g. expression plasmids, shRNA vectors) and RNA (e.g. siRNA duplexes) can be transfected using the same transfection protocol. Efficiencies and viabilities achieved with the 96-well system are in the same range as the single-cuvette Nucleofector, i.e. up to 99% transfection efficiency with siRNA duplexes and up to 95% efficiency with plasmid DNA.
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Figure 4: siRNA-mediated depletion of vimentin-mRNA in Jurkat cells Jurkat E6-1 (ATCC TIB 152) (200,000 cells) were transfected with a siRNA duplex directed against endogenous vimentin. 24h post transfection vimentin mRNA levels were analyzed by RealTime PCR. Relative expressions compared to untreated control sample (well 31, set to 100%) are shown. (External data kindly provided by C. Merz, Schering AG, Berlin)
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High reproducibility
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Jurkat cells (ATCC) were transfected with pmaxGFP. The results shown reflect not only the outstanding data quality but also the absence of technical artifacts such as plate- or edge effects. The standard deviation was determined to 1.04% for all samples, demonstrating an excellent reproducibility.
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Figure 5: Example for intra-plate reproducibilityJurkat cells (ATCC) were transfected with pmaxGFP. Analysis was performed by flow cytometry 24h post nucleofection. The transfection efficiency of each well is shown per column of a 96-well Nucleocuvette Module. Column 4 contained two control samples (no pulse, no plasmid). The standard deviations for the single columns were 1.26 / 0.95 / 0.88 / 0.94% with an over all standard deviation of 1.04% for all samples. (External data kindly provided by C. Merz, Schering AG, Berlin)
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Ordering Information
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| Cat.No. | Devices | | AAD-1001 | Nucleofector II Device | | AAM-1001 | 96-well Shuttle
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| Cat.No. | Cell Line 96-well Nucleofector Kits with optimized protocols for cell lines | | VHCA-1001 | Cell Line 96-well Nucleofector Kit SE (96 reactions) | | VHCA-2001 | Cell Line 96-well Nucleofector Kit SE (960 reactions) | | VHCA-1002 | Cell Line 96-well Nucleofector Kit SF (96 reactions) | | VHCA-2002 | Cell Line 96-well Nucleofector Kit SF (960 reactions) | | VHCA-1003 | Cell Line 96-well Nucleofector Kit SG (96 reactions) | | VHCA-2003 | Cell Line 96-well Nucleofector Kit SG (960 reactions) | | | | | VHCO-1001 | 96-well Cell Line Optimization Kit | | | | | Cat.No. | Cell-type specific 96-well Nucleofector Kits for primary cells | | VHPA-1001 | Human B Cell (96 reactions) | | VHPA-2001 | Human B Cell (960 reactions) | | VHPA-1002 | Human T Cell (96 reactions) | | VHPA-2002 | Human T Cell (960 reactions) | | VHPA-1006 | Mouse T Cell (96 reactions) | | VHPA-2006 | Mouse T Cell (960 reactions) | | VHPA-1007 | Human Monocyte (96 reactions) | | VHPA-2007 | Human Monocyte (960 reactions) | | VHPB-1002 | HUVEC (96 reactions) | | VHPB-2002 | HUVEC (960 reactions) | | VHPD-1001 | NHDF (96 reactions) | | VHPD-2001 | NHDF (960 reactions) | | VHPI-1003 | Neurons, basic (96 reactions) | | VHPI-2003 | Neurons, basci (960 reactions) |
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96-well Shuttle includes Laptop with Nucleofector 96-well Shuttle Software.
Kits for 96/960 reactions contain Nucleofector Solution, Supplement, pmaxGFP plasmid as positive control and 1/10 Nucleocuvette Plate(s).
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Nucleofector 96-well Shuttle |
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The Nucleofector Technology, comprising Nucleofection Process, Nucleofector Device, Nucleofector Solutions, Nucleofector 96-well Shuttle System and 96-well Nucleocuvette plates and modules are covered by patent and/or patent-pending rights owned by amaxa.
amaxa, Nucleofector, Nucleofection, Nucleocuvette, 96-well Shuttle and maxGFP are trademarks of amaxa AG.
ATCC and the ATCC Catalog Marks are trademarks of ATCC used under License.
Other product and company names mentioned herein are the trademarks of their respective owners.
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