They are resistant to easy transfection methods and lately we have been having issues with our outdated retroviral system. The amaxa nucleofector technology is a transfection system designed to transfer genes into cells quickly and efficiently. The nucleofector ii device is protected by two main fuses. To our knowledge, systematic and efficient methods to transfect human induced pluripotent stem cells ipscs have not been reported. Amaxa biosystems has a new electroporationbased transfection method that enables the dna to directly enter the nucleus while cells are. Indeed, as for regular electroporation methods, poor cell viability has been a major. Ingenio electroporation solution competitive comparison. These are combined with different, cell type specific reagents. An overview of improved electroporation via nucleofector technology and the. The nucleofectortm 2b device or older generations i or ii can also be used for bacteria transformation by using alternative cuvettes. For plasmid transfections, the human stem cell nucleofector kit 1 solution was used on the amaxa nucleofector 2b device program a23. Oct 27, 2009 cells were suspended in nucleofector solution to a final concentration of 2. High efficiency electroporation of hard to transfect cell lines, stem cells and. An efficient method for electroporation of small interfering rnas into.
For nucleofection we used the amaxa basic nucleofector kit for primary. Amaxa nucleofector ii electroporation machine lonza more information. Easy and effective procedure of nucleofector technology by lonza, usa for more information, please visit. Nucleofector technology drives research in numerous different. Comparison of nucleofector ii and 4d nucleofector devices with. The tpm3ntrk1 rearrangement is a recurring event in. It allows efficient transfection of hardtotransfect cell lines and primary cells with different substrates, e. The bottom line is that the nucleofector ii is the right choice for fast, easy and efficient transfection of hard to transfect cells. A nucleofectortm device that comprises unique electrical parameters pre programmed for each optimized cell type, to deliver the substrate directly into the cell.
May 04, 2004 the amaxa nucleofector technology is a transfection system designed to transfer genes into cells quickly and efficiently. Jurkat cells dsmz acc 282 were transfected with the cell line nucleofector kit v, program x001 and 2 g of. Based on electroporation, nucleofection utilizes novel and unique electrical parameters generated by a special nucleofector device. Please note that the amaxa nucleofector technology is not intended to be used for diagnostic purposes or for testing or treatment in humans the nucleofector technology, comprising nucleofection process, nucleofector device, nucleofector solutions, nucleofector. Enhanced nucleic acid delivery using conventional electroporation devices that provide. Despite recent advancements in genetic modification, transfection of human embryonic stem cells hescs remains a capricious process. Trouble with amaxa nucleofection in my lab we need to deliver dna to b cells. Stable strains of il3 independent baf3 cells were established by posttransfection selection of cells for two weeks with 800. The nucleofectortm 2b device is the basic cuvette based transfection system for transfection of hardtotransfect cell lines and primary cells.
Nucleofector ii from amaxa biosystems biocompare product. Use the device with lonzas certified nucleofector solutions and. The nucleofector device is essentially a cell electroporator that is programmed with various profiles that are defined electrical parameters for specific cell types. The nucleofector ii2b device can also be used for bacteria transformation by using the special deisgned electroporation cuvettes for bacteria.
The nucleofector kits nucleofector kits contain a specifi c nucleofector solution and supplement, specifi ed cuvettes, pipettes, and the pmaxgfp control vector. The components of the amaxa nucleofector technology. A new, independent system supporting up to 384 transfections in parallel. Perform the nucleofection as per manufacturers instruction using an amaxa nucleofector ii device.
The nucleofector tm 2b device or older generations i or ii can also be used for bacteria transformation by using alternative cuvettes. In these optimized protocols the best nucleofection conditions. The cells were transfected using either 30 n m or 300 n m of sirna or 2. Nucleofection is a valuable transfection method for. Amaxa 4dnucleofector optimization protocol for cell lines. Both are inside a receptacle incorporated in the inner power socket see figure 2, page 7. This means that the user does not need to optimize the pulse parameters manually saving a lot of time, cells and reagents. Dna vectors or sirna oligonucleotides, in low throughput format. Eukaryotic system biorad and in the amaxa nucleofector ii 2b device lonza. Nucleofector supplement is added to the nucleofector solution it is stable for three months at 4c. The amaxa nucleofector technology notebook control unit. Disconnect the nucleofector ii device from power and insert a small flat screwdriver into the slot on the righthand side of the. Jul 18, 2017 the nucleofector technology offers high flexibility with respect to applications, since the same transfection parameters apply for any nucleic acid substrate used. The ingenio electroporation kit was used to transfect 2 x 10 6 ips cells on the amaxa nucleofector ii device.
They are designed to connect amaxa certified addon devices to the. May 25, 2007 amaxa s website also provides you with detail information and a list of easy to follow optimized protocols for a variety of cell types. For instance, the squarewave pulsebased new electroporation devices, such as lonza nucleofector ii electroporation system. Transfer the cells with dna to a cuvette supplied with the nucleofector kit. Amaxa nucleofector ii electroporation machine lonza uk. Nucleofector kits are only functional in combination with the nucleofector device. Validation of cytokines for the chemically defined manufacturing of human cd34 positive hematopoietic precursor cells. Insert screw driver supplied with the 4dnucleofector system in the lower hole of.
In case of a blown fuse, you can easily replace it. The pcdna expression vectors containing trka, trkb and trkc were introduced into the baf3 cells by electroporation amaxa nucleofector ii device. A system enabling transfection of cells by nucleofection in several formats. Only specialized programs for bacterial transformation. The nucleofector technology offers high flexibility with respect to applications, since the same transfection parameters apply for any nucleic acid substrate used. In amaxa certified cuvettes, the cells were nucleofected by using the program y. Amaxa nucleofector ii electroporation machine lonza. Oct 26, 2010 easy and effective procedure of nucleofector technology by lonza, usa for more information, please visit.
The first is the nucleofector tm device, which offers a unique portfolio of electrical parameters, displayed as distinct programs that are adapted to the requirements of specific cell types. The nucleofector ii device is the basic device for single cuvette applications. They are not compatible with the nucleofector ii 2b device. A012, a0, a023, a027 and b016 a12, a, a23, a27 and b16. A nucleofectortm device that comprises unique electrical parameters. There are cell typespecific optimized protocols or recommendations available in our knowledge database. In these optimized protocols the best nucleofection conditions are indicated. Two electroporators were used with different electroporation kits. Transfecting and nucleofecting human induced pluripotent. Easy and effective procedure of nucleofector technology by lonza, usa. Cells were electroporated in parallel with an egfp reporter vector using a nucleofector ii device amaxa and either the ingenio electroporation solution or the nucleofector kit v amaxa according to amaxas recommended protocol for each cell line hl60 t019. With the amaxa nucleofector technology primary cells and stem cells, as well as cell lines, can be. Cells were analyzed 24 hours post nucleofection using light a and fluorescence microscopy b. Technology, originally introduced into the market by legacy amaxa in 2001.
Amaxa nucleofector ii electroporation machine lonza amaxaa nucleofectora technology is a highly efficient nonviral method for transfection. Ht nucleofector high throughput nucleofection system. Cells were electroporated with 8 g zsgreen expressing plasmid clontech in 100 l and plated in 6well plates at 0. The correspond,biological,biology supply,biology supplies. Cells were electroporated by use of the amaxa nucleofector ii instrument with 0. Amaxa nucleofector technology is a highly efficient nonviral method for transfection. Supplement is added to the nucleofector solution, it is stable for three months at 4 c.
A very precise and tunable system fot silencing specific alleles in ipscs is described. An efficient low cost method for gene transfer to t. This means that the user does not need to optimize the pulse parameters manually saving a. These contain lonza certified cuvettes, pipettes, nucleofector solution and supplement. Cells were resuspended in solution from nucleofector kit r, following the amaxa guidelines for cell line transfection. Ingenio electroporation solution and kits mirus bio llc. Amaxa biosystems nucleofector ii amaxa nucleofector technology is a highly efficient nonviral method for transfection. The nucleofector ii 2b transfection device is the single cuvette based system that has been used in research labs since 2001, and formerly known as the amaxa device. We herein report a convenient and affordable method based on in house made buffers, generic cuvettes and utilization of the widely available lonza nucleofector ii device to promote efficient gene transfer to t lymphocytes. Biopharmaceuticals are one of the key growth drivers of the pharmaceutical and biotechnology industries. Nucleofector 2b manual for research only the lonza picturepark.
Amaxa s website also provides you with detail information and a list of easy to follow optimized protocols for a variety of cell types. Nucleofector ii device from lonza group ltd selectscience. Cell line nucleofector kit v from lonza cologne ag,for nucleofection of optimized cell lines in combination with the corresponding optimized protocol including the nucleofection parameters. Email to friends share on facebook opens in a new window or tab share on twitter. The nucleofectortm 2b device is a successor of the original platform that was introduced. Electroporation, transfection and nucleofection researchgate, the professional. The major drawbacks of nucleofection are the cost the nucleofector machine and specific kits containing buffer and cuvettes are required and the induced cell mortality. A comparative study of transfection methods for rna. Ppt 4dnucleofector powerpoint presentation free to. Efficient transfection of primary cells cell lines. The nucleofector ii2b device can also be used for bacteria transformation by using the. In comparison to the nucleofector ii device the viability augmented from 27.
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