Smartseq v4 ultra low input rna kit for the fluidigm c1 system, ifcs user manual 032416. Each tet on 3g inducible gene expression system includes two core elements. Lentix teton 3g inducible expression system user manual. Clontech offers many gfp products, including expression vectors, sequencing primers, purified recombinant gfp proteins, and gfpspecific monoclonal and polyclonal antibodies. The user friendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning. Does anyone have a problem using clontech s hd infusion. Bd genomewalker kits user manual university of north. Both systems offer inducible expression of your gene of interest based on the presence or absence of dox andor tet. See appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. Teton 3g is the gold standard tetracycline inducible expression system for. Safety guidelines for working with recombinant aav. Pr9z3431 3 infusion advantage pcr cloning kit user manual infusion advantage pcr cloning kits are designed to join multiple pieces of dna that have. Clontech is required by its licensing agreement to submit a report of all purchasers of the tet controllable expression system to tet systems. Aavpro tet one inducible expression system aav2 user manual b.
User manual tetoff and teton gene expression systems user manual united statescanada 800. The precise reason for the upper limit on bd genomewalker products is not clear. Knockout rnai systems user manual dartmouth college. The clontech infusion pcr cloning system allows you to fuse of the ends of the pcr fragment to the homologous ends of a linearized vector. Tre tight is the name of the second generation clontech promoter which can be induced by the tta regulator in the tet off system, or it can be induced by the rtta regulator in the tet on system depending on your cloning needs. Please refer to your matchmaker system specific user manual for further information on library screening strategies and specific protocols. Any suggestions for establishing a tetracycline inducible. Tetoff and teton gene expression systems user manual. Ptre3g retains very strong induced expression when activated by transactivator binding. Before the tetone systems were developed, clontechs teton and tetoff. When expressed in either eukaryotic or prokaryotic cells and illuminated by blue or uv light, gfp yields a bright green fluorescence.
Teton 3g inducible expression systems user manual teton 3g inducible expression. You can eliminate tc contamination problems by using clontech tet system approved fbs. Please look at the manual associated with this plasmid for detailed protocols. Invitrogen gateway recombination cloning technology circumvents traditional restriction enzyme based cloning limitations, enabling you to access virtually any expression system in just a few simple steps. Lentix tetone inducible expression systems user manual. Clontech does not support or recommend this product. Purchase of any clontech twohybrid reagent does not imply or convey a license to practice the twohybrid system covered by these patents. Please refer to your matchmaker systemspecific user manual for further information on library screening strategies and specific protocols. Summary thelentix tet on advanced inducible expression system uses a highly efficient, lentiviralbased gene transfer technology to deliver our tightly regulated and highly responsive tet on advanced gene. Tetone inducible expression system user manual 032315. Also available from clontech are several vectors encoding gfp variants that were developed to improve the use of gfp as a reporter for gene expression and protein. Lentix teton advanced inducible expression system user. Fluidigm and takara bio subsidiary clontech laboratories have jointly launched the smartseq v4 ultra low input rna kit for the fluidigm c1 system.
Clontechs smartseq v4 ultra low input rna kit for the. Teton 3g inducible expression systems user manual pt51481. The teton 3g systems are inducible gene expression systems for mammalian cells. The 3 and 5 regions of homology are generated by adding 15 bp extensions to both pcr primers that precisely match the ends of the linearized vector. The allinone system for the inducible expression of. Tetracycline inducible expression systems teton 3g clontech. Applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material.
The inducible ptre3g promoter, which provides the lowest basal expression of any tetracyclineinducible promoter in the absence of bound transactivator. Pr782344 lentix teton advanced inducible expression system user manual e. The allinone system for the inducible expression of shrna. The protocols in this manual require the handling of aav vectors. Infusion advantage protocol overview the table below is a general outline of the protocol used in the infusion advantage pcr cloning kits.
Introduction the creator dna cloning kits provide a revolutionary system for transferring a target gene directly into multiple expression vectors, without the need for timeconsuming subcloning steps. Tet on 3g inducible expression systems user manual pt51481. Introduction continued in the bd genomewalker protocol, the use of ld pcr extends the range of possible pcr products to about 6 kb. Adenoviral modulation of the tumorassociated system l. This serum has been functionally tested in the tet systems to ensure against possible tc contamination.
Introduction continued see appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. Tet off and tet on gene expression systems user manual. The tetone systems are inducible gene expression systems for mammalian cells that contain all the necessary components in a single plasmid, lentiviral, or. Tetone inducible expression system user manual takara bio. Tetracycline inducible expressiontetone systems clontech. The rapid titer method uses an adenovirusspecific antibody to identify infected cells. Introduction continued optimized vectors facilitate downstream confirmation the matchmaker system 3 dnabd and ad fusion vectors, pgbkt7 and pgadt7, were designed. Schematic of gene regulation in the bd tetoff and teton systems. Apr 06, 2000 applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material.
The kit is available for four and 10 integrated fluidic circuits, and allows for highquality cdna synthesis starting from 96 single cells that have been isolated and processed with the c1 system. Pr63158 knockout rnai systems user manual that is complementary to a particular mrna, to suppress the expression of the par1 gene in c. Clontechs smart cdna synthesis kit biocompare product. Target cells that express the teton 3g transactivator protein and contain a. Adenoviral modulation of the tumorassociated system l amino. Selecting cut sites and copying the sequence will also activate enzymes. Teton 3g inducible expression systems user manual takara bio. Matchmakergold yeast twohybrid system user manual pt40841 pr033493 cat.
Aavpro tetone inducible expression system aav2 user manual. Plasmid resources, addgene 12 cumateinducible systems. Teton advanced inducible gene expression systems user manual. Tet one inducible expression system user manual 032315. It is imperative to fully understand the potential hazards of and necessary precautions for laboratory use of these vectors. Lentix teton advanced inducible expression system user manual. Does anyone have a problem using clontech s hd infusion cloning system. Novartis developmental and molecular pathways, cambridge, ma, usa.
Convert pcr primers into infusion primers infusion adaptors, a vectorspecific sequences of approximately 15 nr long, will be added on the 5end of your primers. List of components the smartseq v4 ultra low input rna kit for the fluidigm c1 system, ifcs consists of the smartseq v4. A system for regulated expression in mammalian cells. Trace amounts of tetracycline in regular fbs might trigger shrna expression. The packaging mix includes an expression vector for the tetoff transcriptional activator tta and uses tet. User manual tet off and tet on gene expression systems user manual. Does anyone have problems with the lentixt htx packaging. Tbusa, formerly known as clontech laboratories, inc. Bd biosciences clontech now offers a faster, more convenient way to titer recombinant adenovirus.
Summary thelentix teton advanced inducible expression system uses a highly efficient, lentiviralbased gene transfer technology to deliver our tightly regulated and highly responsive teton advanced gene. Enter i sequences of pcr primers 1 and 2, ii fragment of vector sequence of about 40 bp with the cloning site in. In this approach, target dna molecules and cloning vector are generated by pcr with 610 bases of homology between the neighboring fragments. Infusion advantage pcr cloning kit user manual protocol no.
A solution to this problem can be found in clontechs smart cdna synthesis kit. For a complete list of tet systems references, visit our web site at. The name was changed to clarify that this plasmid does not contain and is not related to the trademarked tetonr system sold by clontech. Pr782344 3 lentix tet on advanced inducible expression system user manual a. Mountain view, ca 94043 technical support us email. It can be used as either a standalone program or a library.
Cut sites of enzymes that you select are highlighted to help guide your work. Novartis plkoteton 1 the allinone system for the inducible expression of shrna dmitri wiederschain, ph. Clontech is required by its licensing agreement to submit a report of all purchasers of the tetcontrollable expression system to tet systems. Tetracycline inducible expression, clontech, 2002 11 tetracycline tet inducible expression. The bd clontech adenox rapid titer kit is a complete antibodybased system that lets you measure adenoviral titer in as little as three days. I am following the standard protocol first transfect with pcdna6. Tre tight is the name of the second generation clontech promoter which can be induced by the tta regulator in the tetoff system, or it can be induced by the rtta regulator in the teton system depending on your cloning needs. Its goal is to generate good quality tetrahedral meshes suitable for numerical methods and scientific computing. The name was changed to clarify that this plasmid does not contain and is not related to the trademarked tet onr system sold by clontech. For the last couple of months i have been trying to establish an inducible hacat cell line using the trex system from invitrogen.
Benefits of the tetadvanced expression systems thetetoff advanced andteton advanced systems yield very high maximal expression levels and extremely. Kg prior to purchasing these reagents or using them for any purpose. Living colors user manual v olume ii stanford university. Clontech, 631187, lentix teton 3g inducible expression system. The userfriendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning.
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