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