The table below shows data for the cell type and nucleofector platform selected. Software updates lonza reserves its rights to change the described software without announcement or to generate updates of the software. Nucleofector technology requires a license from amaxa gmbh. The optimization strategy is suitable both for optimizing nucleofection of plasmid dna, as well as, sirna oligonucleotides. Sep 24, 2015 preparation get a boneless pork shoulder butt or boneless pork loin. Does anyone have experience with amaxa nucleofector based. Nucleofector 2b manual for research only the lonza picturepark. Nucleofector technology stretching transfection dimensions. Tradein your old nucleofector device nucleofector i or ii until 30 june 2014 and save at least 2000 gbp when purchasing a new 4d nucleofector system. Optimizing these methods is possible, but many variables have to be taken into account. Download document genome editing using nucleofector tm technology technical reference guide providing a brief introduction to genome editing tools and technical tips for zfn, talen or crispr delivery using the nucleofector tm technology. Nevertheless, we developed in house methods for ecm 630 electroporator machine.
Successful genome editing using zfn, talen or crispr requires efficient delivery of such tools into the cell type of interest. I tried 5 nucleofection programs, 2 buffers and several different cell densities. For all endotoxin detection assays please enter the kit product code and lot number from the kit box label. By continuing to browse the site, you are agreeing to our use of cookies. The nucleofector tm technology uses a specific combination of optimized electrical parameters and cell typespecific solutions which enables transfer of a molecule directly into the cells nucleus. The controlling unit of the 4d nucleofector tm system is suited to handle various fuctional units for efficient transfection of hardtotransfect cell lines and primary cells in different formats. Ingenio electroporation products for high efficiency electroporation. Wolmanized heavy duty wood is used in a variety of highway applications including. Pc12 cells were transfected with the cell line nucleofector kit v, program u029 and 2 g of pmaxgfp vector. It includes 4dnucleofector system, 96well shuttle system, ht nucleofector system, nucleofector 2b device. Having tested various fibroblastoid cell types, high transfection efficiencies could be achieved using one of the programs indicated below. Electroporation by nucleofector is the best nonviral. H negative control no program negative control no program required material note please make sure that the entire supplement is added to the nucleofector solution. Turn on the nucleofector by using the power switch on the rear side.
Virusfree induction of pluripotency and subsequent excision of reprogramming factors. Longterm protection plus advantageous environmental, economical, and handling features make aczatreated crossties an exceptional choice for railroads life cycle assessment. General protocol for nucleofection of adherent cell lines. Cells were analyzed 24 hours post nucleofection using light a and fluorescence microscopy b. Nucleofector tm 2b device the basic, single cuvette based nucleofector tm system for efficient transfection of hardtotransfect cell lines and primary cells with different substrates e. Once the nucleofector supplement is added to the nucleofector solution it is stable for three months at 4c. Select nucleofector program insert cuvette press start button x. Download white paper we use cookies to deliver the best service to you. It includes 4d nucleofector system, 96well shuttle system, ht nucleofector system, nucleofector. The ht nucleofector software is preinstalled on a laptop computer.
Nucleofector technology offers high flexibility within applications, since the same. The nucleofector device from lonza group ltd selectscience. Therapeutic cell culture media, research tools, testing solutions and quality control software. The 4d nucleofector tm x unit is also required in order to run the 96well shuttle tm device there are cell typespecific optimized protocols or recommendations available in our knowledge database. For preparing aliquots, mix nucleofector solution and supplement in a ratio of 4. They are based on cell culture methods, where live cells are grown in vitro. Lonza leads the innovation for endotoxin detection products, services and software. The nucleofector programs available correspond to the list of optimized. Either way, they are all made from whole or cut, boneless pork loin preferably with a nice fat cap intact. A further set of programs is tested based on the three best nucleofector programs and the best nucleofector solution from the initial experiment. Aug 10, 2015 lonzas nucleofector technology is an improved electroporation technology that can help you achieve transfection results never before possible in primary cells, stem cells and hard to. Cell culture assays provide a means of quantitatively analyzing the presence, amount, or functional activity.
Amaxa 4dnucleofector optimization protocol for cell lines. The electrical settings are preprogrammed for each optimized cell type and can be selected via the device or pc software. They are based on cell culture methods, where live cells are grown in vitro and used as model systems to assess the biochemistry and physiology of both healthy and diseased cells. Amaxa 96well shuttle protocol for primary cell optimization. Use of lonzas nucleofector technology andor related software requires a license from lonza cologne gmbh. For determination of the optimal transfection conditions for any difficulttotransfect cell line that lacks an optimized protocol for the nucleofector tm iii2b device. The duration is also dependent on how long the nucleofector device has been turned on as well as the number. With lonzas researchuse human bone marrow and peripheral blood immune cells you dont have to hassle with finding donors and isolating specific cell types, because purified, high viability cells are shipped directly to you.
Cured pork loin is commonly known as lonza loin but may also be referred to by some as lomo in spanish or lombo. Endotoxin detection is a part of lonza biosciences qc testing and analytical services that offers a comprehensive qc platform view the catalog and learn more about lonza s endotoxin products and services. Software updates will be announced by email and provided on a cd or as download. Purchasers are granted a nonexclusive, nontransferable license for the use of amaxas nucleofector technology for research and development purposes, the terms of which are disclosed in detail in the license agreement accompanying the shipped nucleofector device. Lonza walkersville 4d nucleofector core unit 4d nucleofector core unitfl1. Ppt 4d nucleofector powerpoint presentation free to download id. If the experimental setup is as desired, load the samples for unit. Lonza s nucleofector technology is an improved electroporation technology that can help you achieve transfection results never before possible in primary cells, stem cells.
Amaxa nucleofector protocol for rat oligodendrocytes. It supports nucleofection of various cell numbers in different formats. We do not just work in the life sciences industries, we are transforming them. Nucleofector supplement is added to the nucleofector solution it is stable for three months at 4c. Please try both programs as transfection efficiency may vary. The offer is valid until 30 june 2014 and only for nucleofector i or ii devices in uk. Manuals for all versions are available for download at. Transfer mixture to cuvette and insert into nucleofector. Only use the software in connection with the nucleofector. Optimized protocol for human stem cells lonza bioscience. Genomic editing, nucleofection, and the creation of nextgeneration cellbased assays. The translational research institute is a medical research facility bringing together the university of queensland, metro south health, queensland institute of technology and mater medical research to translate research findings into improved healthcare and increased income and jobs for australia. Learn more about how delivery of crispr plasmids compares with delivery of ribonucleoproteins rnps from our most recent white paper, efficient crisprcas9 delivery using nucleofector. For further technical assistance on using the nucleofector tm technology for crisprbased genome editing, contact lonza scientific support.
The nucleofector technology is a highly efficient nonviral method for transfection. What if i knew how well delivery of crisprcas9 ribonucleoproteins works. Download the free poster as a highresolution pdf, and request your. The primary cell optimization protocol enables you to optimize 96well shuttle conditions for a primary cell of your choice using our primary cell optimization 96well nucleofector kit. Lonza s nonviral nucleofector tm technology has been shown to work as a reliable and sufficient method for cotransferring the required dna or rnabased components into hardtotransfect cell types. Use of lonza s nucleofector technology andor related software requires a license from lonza cologne gmbh. The nucleofector tm technology uses a specific combination of optimized electrical parameters and cell typespecific solutions which enables transfer of a molecule directly into the cells. A free powerpoint ppt presentation displayed as a flash slide show on id.
Cellbased assays and analysis are vital experimental tools in life science research and biomanufacturing. Product overview the 4d nucleofector tm x unit is one of the three functional modules of the 4d nucleofector tm system. Raft system allows longterm culture maintenance and formation of multilayer models. Ppt 4dnucleofector powerpoint presentation free to. Celltech reserved a fixed annual manufacturing capacity at lonza biotecs 1,500l and 15,000l fermenter systems for recombinant microbial products in visp, switzerland, covering the period 2004 to 2010. Transfection using the nucleofector is a patented commercial technique requiring special buffers and programs, the constituents of which are a secret.
Lonza biologics inc offers wide range of pharma products and services which includes nucleofector devices. Make sure that each potential user reads and understands it. Lonza 4dnucleofector nucleofector used miscellaneous. Amaxa cell line optimization nucleofector kit lonza bioscience. Those data are either based on lonza optimized protocols or on results shared from customers who performed an optimization based on our guidelines. We have tried dk100 and dn107 programs, however, the postelectroporation viability of. The controlling unit of the 4d nucleofector tm system is suited to handle various fuctional units for efficient transfection of hard to transfect cell lines and primary cells in different formats. Efficient genome editing using nucleofector technology.
Environmental chemonite treated wood shares many of the environmental. The nucleofection condition with the highest efficiency and lowest mortality is selected for all subsequent experiments and can be also used in the 100 l single nucleocuvette. Approved agricultural applications for wolmanized heavy duty wood include timbers, poles, piles, and fence posts used as structural members on farms, as well as plywood, permanent wood foundations, and gluelaminated timbers. We do not just work in the life sciences industries, we are. Market trends support all four areas in which lonza bioscience operates. To speak with a representative, please fill out the contact form below or call us directly. Each solution and supplement is individually developed for every primary cell type. When done, itll be about 1 34 lb 2 roll the pork in the salt rub. The ratio of nucleofector solution to supplement is 4. The nucleofector technology relies on the combination of a nucleofector device and cell specifi c nucleofector kits. The adobe flash plugin is needed to view this content. All content is posted anonymously by employees working at lonza. The duration time of a program is dependent on the number of the program. The nucleofector device delivers unique electrical parameters.
Due to this independency from cell proliferation, it allows for efficient transfection of even nondividing primary cells like resting t cells or neurons and also speeds up expression. Virusfree induction of pluripotency and subsequent excision. Railroad crossties utility poles marine agricultural railroad crossties chemonite acza is made for heavy duty applications. Cells were then electroporated using a nucleofector lonza set to program d032. Purchasers are granted a nonexclusive, nontransferable. Lonza 4d nucleofector core unit w x and y unit nucleofector. Please enter all of the catalog and lot numbers, including all zeros, located on the product label. Product overview the cell line optimization 4d nucleofector tm x kit is the ideal tool to conveniently and rapidly determine nucleofection conditions for cell lines lacking an optimized protocol for the 4d nucleofector tm x unit with the unique capability of the 4d nucleofector tm x unit to address individual wells of a 16well nucleocuvette tm strip with different programs. The nucleofection condition with the highest efficiency and lowest mortality is selected for. Lonza nucleofector technology at tri translational research. Importance of hepatocytes and other primary cells for in vitro admetox studies in drug discovery and development. Only use the software once you have read and understood the nucleofector 96well shuttle software manual. Lonza was to provide technology transfer, scaleup, cgmp manufacturing and quality control testing services for celltech.
Optimized protocols that are provided with the nucleofector kits which. A ed158 no program b eh158 ff158 c eh166 fp150 d er7 if100 no program addition of nucleofector solution and pmaxgfp vector, but no application of program the nucleofector program. Lonza walkersville p3 primary cell 4dnucleofector x kit l. Protocols that are provided with the nucleofector kits which can be downloaded at. It belongs to bioresearch transfection services and produtcs category. All electroporations were performed on the lonza nucleofector 2b program u014 for hspcs and t cells. Mef nucleofector kit 2 amaxa and program t20 were used for nucleofection according to the manufacturer. Use of lonza s nucleofector technology andor related software requires a license from lonza. Ingenio cuvettes for the ezporator electroporation system and lonzaamaxa nucleofector ii2b devices ingenio cuvettes for the ezporator electroporation system and all conventional electroporators such as biorad and harvardbtx ingenio cell droppers.
For longterm storage, pmaxgfp vector is ideally stored at 20c. We have tried dk100 and dn107 programs, however, the postelectroporation. Those data are either based on lonza optimized protocols or on results shared from customers who. We have a lonza nucleofector and it seems that this is the best way to get dna in those cells.
In case no data are shown for the selected nucleofector. The keypads are active when the power indicator 1 is on. The expiration date is printed on the solution box. To access a certificate of analysis, please enter the catalog number and lot number located on the product label. The p3 primary cell 4dnucleofector x kit l is one of five 4dnucleofector kits suited for transfecting primary cells in the 100 l nucleocuvettetm vessel format when working with the 4dnucleofectortm x unit. What we do today is helping to create a healthier, happier world tomorrow. Does anyone have any experience with transfection of raw 246. In our hands, nucleofector program g worked well for both embryonic rat neurons as well as postnatal mouse neurons programs recommended by lonza are o005 for mouse neurons and o003 or g for rat neurons. Genomic editing, nucleofection, and the creation of next.
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