Clontech tet system user manual

Teton 3g inducible expression systems user manual teton 3g inducible expression. The allinone system for the inducible expression of. Safety guidelines for working with recombinant aav. Knockout rnai systems user manual dartmouth college. Mirx inducible mirna systems user manual takara bio. I am following the standard protocol first transfect with pcdna6. Cut sites of enzymes that you select are highlighted to help guide your work. Tet off and tet on gene expression systems user manual. 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. It is imperative to fully understand the potential hazards of and necessary precautions for laboratory use of these vectors.

A system for regulated expression in mammalian cells. See appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. Clontechs smart cdna synthesis kit biocompare product. Its goal is to generate good quality tetrahedral meshes suitable for numerical methods and scientific computing. User manual tetoff and teton gene expression systems user manual united statescanada 800. It can be used as either a standalone program or a library. Selecting cut sites and copying the sequence will also activate enzymes.

Smartseq v4 ultra low input rna kit for the fluidigm c1 system, ifcs user manual 032416. Lentix tetone inducible expression systems user manual. Introduction continued in the bd genomewalker protocol, the use of ld pcr extends the range of possible pcr products to about 6 kb. Does anyone have a problem using clontech s hd infusion cloning system. In this approach, target dna molecules and cloning vector are generated by pcr with 610 bases of homology between the neighboring fragments. For the last couple of months i have been trying to establish an inducible hacat cell line using the trex system from invitrogen. Lentix teton advanced inducible expression system user manual. 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. Does anyone have a problem using clontech s hd infusion. The packaging mix includes an expression vector for the tetoff transcriptional activator tta and uses tet. The tetone systems are inducible gene expression systems for mammalian cells that contain all the necessary components in a single plasmid, lentiviral, or. Plasmid resources, addgene 12 cumateinducible systems. Please look at the manual associated with this plasmid for detailed protocols. The bd clontech adenox rapid titer kit is a complete antibodybased system that lets you measure adenoviral titer in as little as three days.

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. Pr63158 knockout rnai systems user manual that is complementary to a particular mrna, to suppress the expression of the par1 gene in c. Fluidigm and takara bio subsidiary clontech laboratories have jointly launched the smartseq v4 ultra low input rna kit for the fluidigm c1 system. Introduction continued optimized vectors facilitate downstream confirmation the matchmaker system 3 dnabd and ad fusion vectors, pgbkt7 and pgadt7, were designed. When expressed in either eukaryotic or prokaryotic cells and illuminated by blue or uv light, gfp yields a bright green fluorescence. Structure and formation of the rcfp fluor ophore the rcfp.

The name was changed to clarify that this plasmid does not contain and is not related to the trademarked tetonr system sold by clontech. Ptre3g retains very strong induced expression when activated by transactivator binding. Does anyone have problems with the lentixt htx packaging. Tbusa, formerly known as clontech laboratories, inc. Clontechs smartseq v4 ultra low input rna kit for the. 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. The rapid titer method uses an adenovirusspecific antibody to identify infected cells. Tet on 3g inducible expression systems user manual pt51481. Tetone inducible expression system user manual 032315. For a complete list of tet systems references, visit our web site at. Clontech is required by its licensing agreement to submit a report of all purchasers of the tet controllable expression system to tet systems. This serum has been functionally tested in the tet systems to ensure against possible tc contamination. Pr782344 3 lentix tet on advanced inducible expression system user manual a. Kg prior to purchasing these reagents or using them for any purpose.

Teton advanced inducible gene expression systems user manual. The user friendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning. Tetracycline inducible expression, clontech, 2002 11 tetracycline tet inducible expression. Bd genomewalker kits user manual university of north. 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. Benefits of the tetadvanced expression systems thetetoff advanced andteton advanced systems yield very high maximal expression levels and extremely. Tetracycline inducible expression systems teton 3g clontech. Pr782344 lentix teton advanced inducible expression system user manual e. Before the tetone systems were developed, clontechs teton and tetoff. Bd biosciences clontech now offers a faster, more convenient way to titer recombinant adenovirus.

The allinone system for the inducible expression of shrna. 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. Applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material. Trace amounts of tetracycline in regular fbs might trigger shrna expression. Clontech, 631187, lentix teton 3g inducible expression system. User manual tet off and tet on gene expression systems user manual. 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. Infusion advantage protocol overview the table below is a general outline of the protocol used in the infusion advantage pcr cloning kits. Teton 3g is the gold standard tetracycline inducible expression system for. Novartis plkoteton 1 the allinone system for the inducible expression of shrna dmitri wiederschain, ph.

Lentix teton 3g inducible expression system user manual. Tetracycline inducible expressiontetone systems clontech. Any suggestions for establishing a tetracycline inducible. Tet one inducible expression system user manual 032315. The teton 3g systems are inducible gene expression systems for mammalian cells. Aavpro tet one inducible expression system aav2 user manual b. Ive followed all of the instructions according to the clontech protocol and im not sure what could be going wrong. Teton 3g inducible expression systems user manual pt51481. Introduction continued see appendix a or the vector information packets provided for maps and detailed information on the tet system vectors. 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.

Infusion advantage pcr cloning kit user manual protocol no. List of components the smartseq v4 ultra low input rna kit for the fluidigm c1 system, ifcs consists of the smartseq v4. Schematic of gene regulation in the bd tetoff and teton systems. Please refer to your matchmaker systemspecific user manual for further information on library screening strategies and specific protocols. Purchase of any clontech twohybrid reagent does not imply or convey a license to practice the twohybrid system covered by these patents. A solution to this problem can be found in clontechs smart cdna synthesis kit. Aavpro tetone inducible expression system aav2 user manual. You can eliminate tc contamination problems by using clontech tet system approved fbs. 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. The userfriendly dna engineering method allows combining multiple pcr fragment assembly, nucleotide sequence alteration and directional cloning. Target cells that express the teton 3g transactivator protein and contain a. Apr 06, 2000 applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material. Lentix teton advanced inducible expression system user.

Novartis developmental and molecular pathways, cambridge, ma, usa. Adenoviral modulation of the tumorassociated system l amino. Tetone inducible expression system user manual takara bio. Both systems offer inducible expression of your gene of interest based on the presence or absence of dox andor tet. Teton 3g inducible expression systems user manual takara bio. 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. Clontech does not support or recommend this product. The precise reason for the upper limit on bd genomewalker products is not clear. The inducible ptre3g promoter, which provides the lowest basal expression of any tetracyclineinducible promoter in the absence of bound transactivator. Summary the human genome project has generated the sequences of thousands of genes aaronson et al. Adenoviral modulation of the tumorassociated system l. Enter i sequences of pcr primers 1 and 2, ii fragment of vector sequence of about 40 bp with the cloning site in.

Tetoff and teton gene expression systems user manual. Mountain view, ca 94043 technical support us email. The protocols in this manual require the handling of aav vectors. 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.