Adenoviral modulation of the tumorassociated system l. Adenoviral modulation of the tumorassociated system l amino. Pr782344 lentix teton advanced inducible expression system user manual e. Enter i sequences of pcr primers 1 and 2, ii fragment of vector sequence of about 40 bp with the cloning site in. Lentix teton 3g inducible expression system user manual. You can eliminate tc contamination problems by using clontech tet system approved fbs. For a complete list of tet systems references, visit our web site at. Selecting cut sites and copying the sequence will also activate enzymes. In this approach, target dna molecules and cloning vector are generated by pcr with 610 bases of homology between the neighboring fragments. Novartis developmental and molecular pathways, cambridge, ma, usa. Teton 3g inducible expression systems user manual pt51481. It can be used as either a standalone program or a library. For the last couple of months i have been trying to establish an inducible hacat cell line using the trex system from invitrogen. Mirx inducible mirna systems user manual takara bio.
Both systems offer inducible expression of your gene of interest based on the presence or absence of dox andor tet. It is imperative to fully understand the potential hazards of and necessary precautions for laboratory use of these vectors. The user friendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning. Bd biosciences clontech now offers a faster, more convenient way to titer recombinant adenovirus. Does anyone have a problem using clontech s hd infusion cloning system. Tet on 3g inducible expression systems user manual pt51481. Mountain view, ca 94043 technical support us email. Tetracycline inducible expression systems teton 3g clontech. A system for regulated expression in mammalian cells. Matchmakergold yeast twohybrid system user manual pt40841 pr033493 cat. 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.
Tetoff and teton gene expression systems user manual. Novartis plkoteton 1 the allinone system for the inducible expression of shrna dmitri wiederschain, ph. Lentix tetone inducible expression systems user manual. Kg prior to purchasing these reagents or using them for any purpose. Tetracycline inducible expression, clontech, 2002 11 tetracycline tet inducible expression.
Pr9z3431 3 infusion advantage pcr cloning kit user manual infusion advantage pcr cloning kits are designed to join multiple pieces of dna that have. The bd clontech adenox rapid titer kit is a complete antibodybased system that lets you measure adenoviral titer in as little as three days. The allinone system for the inducible expression of. The userfriendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning. Its goal is to generate good quality tetrahedral meshes suitable for numerical methods and scientific computing. Tet off and tet on gene expression systems user manual. Does anyone have problems with the lentixt htx packaging system from clontech. Clontech does not support or recommend this product. 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. Bd genomewalker kits user manual university of north. The precise reason for the upper limit on bd genomewalker products is not clear. The rapid titer method uses an adenovirusspecific antibody to identify infected cells. Cut sites of enzymes that you select are highlighted to help guide your work. When expressed in either eukaryotic or prokaryotic cells and illuminated by blue or uv light, gfp yields a bright green fluorescence.
The protocols in this manual require the handling of aav vectors. Summary the human genome project has generated the sequences of thousands of genes aaronson et al. Pr63158 knockout rnai systems user manual that is complementary to a particular mrna, to suppress the expression of the par1 gene in c. List of components the smartseq v4 ultra low input rna kit for. Teton 3g inducible expression systems user manual takara bio. See appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. Structure and formation of the rcfp fluor ophore the rcfp. 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. Safety guidelines for working with recombinant aav. Schematic of gene regulation in the bd tetoff and teton systems. Ptre3g retains very strong induced expression when activated by transactivator binding.
Lentix teton advanced inducible expression system user. I am following the standard protocol first transfect with pcdna6. 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. User manual tet off and tet on gene expression systems user manual. Pr782344 3 lentix tet on advanced inducible expression system user manual a. Infusion advantage protocol overview the table below is a general outline of the protocol used in the infusion advantage pcr cloning kits. Lentix teton advanced inducible expression system user manual. Clontech is required by its licensing agreement to submit a report of all purchasers of the tetcontrollable expression system to tet systems. Clontech, 631187, lentix teton 3g inducible expression system. Trace amounts of tetracycline in regular fbs might trigger shrna expression. Plasmid resources, addgene 12 cumateinducible systems. Please refer to your matchmaker system specific user manual for further information on library screening strategies and specific protocols. Living colors user manual v olume ii stanford university.
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. Aavpro tet one inducible expression system aav2 user manual b. This serum has been functionally tested in the tet systems to ensure against possible tc contamination. A solution to this problem can be found in clontechs smart cdna synthesis kit. Introduction continued optimized vectors facilitate downstream confirmation the matchmaker system 3 dnabd and ad fusion vectors, pgbkt7 and pgadt7, were designed. Tetracycline inducible expressiontetone systems clontech. 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. The inducible ptre3g promoter, which provides the lowest basal expression of any tetracyclineinducible promoter in the absence of bound transactivator. Introduction continued in the bd genomewalker protocol, the use of ld pcr extends the range of possible pcr products to about 6 kb. Tet one inducible expression system user manual 032315. Each tet on 3g inducible gene expression system includes two core elements. Teton 3g is the gold standard tetracycline inducible expression system for. Before the tetone systems were developed, clontechs teton and tetoff.
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. 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. Apr 06, 2000 applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material. Aavpro tetone inducible expression system aav2 user manual. Tbusa, formerly known as clontech laboratories, inc. List of components the smartseq v4 ultra low input rna kit for the fluidigm c1 system, ifcs consists of the smartseq v4. Does anyone have a problem using clontech s hd infusion. User manual tetoff and teton gene expression systems user manual united statescanada 800. The teton 3g systems are inducible gene expression systems for mammalian cells.
Clontech offers many gfp products, including expression vectors, sequencing primers, purified recombinant gfp proteins, and gfpspecific monoclonal and polyclonal antibodies. Tetone inducible expression system user manual 032315. 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. Introduction continued see appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. Purchase of any clontech twohybrid reagent does not imply or convey a license to practice the twohybrid system covered by these patents. The packaging mix includes an expression vector for the tetoff transcriptional activator tta and uses tet. Clontechs smartseq v4 ultra low input rna kit for the. Teton advanced inducible gene expression systems user manual. Does anyone have problems with the lentixt htx packaging. Target cells that express the teton 3g transactivator protein and contain a. To release a highly sensitive kit for human tcell receptor repertoire analysis read this article along with other careers information, tips and advice on biospace. 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.
Please refer to your matchmaker systemspecific user manual for further information on library screening strategies and specific protocols. The name was changed to clarify that this plasmid does not contain and is not related to the trademarked tetonr system sold by clontech. Benefits of the tetadvanced expression systems thetetoff advanced andteton advanced systems yield very high maximal expression levels and extremely. Teton 3g inducible expression systems user manual teton 3g inducible expression. Infusion advantage pcr cloning kit user manual protocol no. Ive followed all of the instructions according to the clontech protocol and im not sure what could be going wrong. Clontechs smart cdna synthesis kit biocompare product. Fluidigm and takara bio subsidiary clontech laboratories have jointly launched the smartseq v4 ultra low input rna kit for the fluidigm c1 system. Clontech is required by its licensing agreement to submit a report of all purchasers of the tet controllable expression system to tet systems. Tetone inducible expression system user manual takara bio.
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