...tetR will bind to tetracycline and not to the TRE, permitting transcription. Tetracycline-dependent promoters...To use tetracycline as a regulator of gene expression, a tetracycline-controlled transactivator (tTA) ...Overview of tetracycline regulated inducible transcriptional activation or repression Plasmid...Collections Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Additional Resources...Tet-responsive Promoters Background Tetracycline Off Tetracycline On Experimental Tips Tet Plasmids Background... with efficient tetracycline uptake. Please search our full site for all tetracycline plasmids. Tet System...19 nucleotide tetracycline operator (tetO) sequence, and is recognized by the tetracycline repressor (tetR...

...Signaling Strains Browse Addgene's collection of synthetic biology related bacterial strains. SynBio Bacterial...Strains for use in Synthetic Biology. Plasmid... Plasmid Collections Synthetic Biology Bacterial Strains Synthetic...Synthetic Biology: Bacterial Strains SynBio Resources SynBio Home Browse All SynBio Assembly Standards SynBio... Bacterial Strains Search the table by keyword or sort by the table headings. Click on the publication...

...Monosynaptic Neuronal Tracing Viral Service: Tools for Monosynaptic Neuronal Tracing These AAV encode tools... EM. 2007b. Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons...encoding tools for monosynpatic retrograde neuronal tracing, to be used with modified rabies virus. Viral...tools for rabies virus-based monosynaptic tracing. These AAV can be used with modified rabies virus to...to perform retrograde neuronal tracing. Note : Addgene does not distribute rabies virus. AAV Vectors for... for Monosynaptic Neuronal Tracing ID Name Description Serotype PI 52473 pAAV-synP-FLEX-splitTVA-EGFP-...Introduction to Rabies Virus-based Monosynaptic Tracing The core idea of the system is to infect the cell...

...AVEXIS (for Avidity-based extracellular interaction screen) in which the extracellular regions of receptor ...EXPRESs Consortium plasmids that encode the extracellular regions of cell surface receptor and secreted... Plasmid Collections EXPRESs Plasmids EXtracellular Plasmid RESource (EXPRESs) Consortium You may...mammalian expression plasmids that encode the extracellular regions of cell surface receptor and secreted...detailed protocol description: Avidity based Extracellular Interaction Screening (AVEXIS) for the scalable...scalable detection of low affinity extracellular receptor-ligand interactions. Kerr SJ and Wright GJ. J. ...9; 480(7378):534-7. Construction of a large extracellular protein interaction network and its resolution...

...CRISPR plasmids that create double strand breaks in DNA....Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...CRISPR nickase plasmids for the generation of single-strand breaks. CRISPR...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants...

... TALengineering Reagents Transcriptional Activators TALE Transcriptional Activator Expression Vectors...methods) can be cloned for expression as TALE transcriptional activators. The Joung Lab has recently shown...plasmids robustly yields highly active artificial transcription factors ( Maeder et al., Nat Methods 2013 )....repeats, and encode either a VP64 or a NF-κB p65 transcriptional activation domain at the carboxy-terminus of...

...for transfecting mammalian cells using linear polyethylenimine. Transfections allow for transient expression...Carefully transfer the transfection mix to the Lenti-X 293T packaging cells. Add the transfection mix dropwise...lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with...protocol to transfect mammalian cells with your plasmid of interest. Protocols... Protocols General Transfection General...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral Service...adapted for different cell lines and different transfection reagents. Last Upload: June 10, 2016 Workflow...

...Introduction Transformation is the process by which foreign DNA is introduced into a cell. Transformation of bacteria...modifications to create bacterial strains that can be more easily transformed and that will help to maintain...prepared for optimal transformation efficiencies upon thawing. For the highest transformation efficiency, we...time when carrying out transformations? If you are not concerned with transformation efficiency (such as ... 20-30 mins) I get very few if any transformants when transforming large plasmids (>10 kb) or BACs, what...Learn how to transform E. coli with your plasmid of interest. Protocols... Protocols Bacterial Transformation Bacterial...

... Protocols ddPCR Titration of AAV Vectors ddPCR Titration of AAV Vectors You...for purified AAV titration. Protocols...You may also like... AAV Titration by qPCR AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular...samples. ITR Master Mix Volume 9X Volume Final Concentration 2X ddPCR Supermix for Probes, no dUTP 10 µL ...the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator....while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled PCR plate. *Pro-Tip... 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft...

...Colony Formation Titration Assay Lentivirus Fluorescence Titration Assay AAV Titration by qPCR Addgene ... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration of Lentivirus...droplets (blue/green). The concentration of RRE positive droplets in the untransduced control should be close...serially diluted and used to transduce HEK293T cells. Genomic DNA is extracted from the target cells and ...+06 7 Untransduced 1 6.4 17940 0.0007134894091 1.43E+02 Table: Example dilutions and titrations Reference...lentivirus vector titration. Protocols...titer range of 1E+05–1E+09 TU/mL, where TU is transducing units and generally represents the number of ...

... to proceed with transfection. Transfect the flask containing 108 mL cells: Transfer 6 mL of BCD TFX into...General Transfection Protocol Recombinant Antibody Purification Protocol Introduction Transfections allow...using Polyethylenimine Max as a transfection reagent. After transfection and expression, the recombinant...Section 2: Transfection Check the cell density and viability: Using a 5 mL pipette, transfer 0.5 mL of ...feeding schedule: Thursday: Transfect cells. Friday (24 h post-transfection): Add 4% BCD Feed. Monday (...post-transfection): Add valproic acid to 3.75 mM, add 4% BCD Feed. Tuesday (120 h post-transfection): Add...Wednesday (144 h post-transfection): Add 4% BCD Feed. Thursday (168 h post-transfection): Harvest antibody...

... Protocols RNA Extraction Without a Kit RNA Extraction Without a Kit...effectively lyse open the cells. Extract RNA from the homogenized sample(s).Transfer tissue/cell lysate to a 4....5 M LiCl solution to bring the concentration of LiCl in the extracted aqueous layer to 2.5 M. Incubate...perform RNA extraction without a kit. Protocols...Reference Introduction This protocol describes how to extract RNA from tissues or cells without a kit. The general...include homogenization/lysis of cells or tissues, extraction of RNA, precipitation, and resuspension. This...

...Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient Ultracentrifugation You may also...medicine, iodixanol can be used as an intravenous isomolar contrast agent. In research, iodixanol is used...the column. We recommend concentrating to a minimum of 500 µl. If the concentrate volume is less than 500...Follow this protocol using Iodixanol gradient ultracentrifugation to get pure adeno-associated virus (AAV) ...Timeline Day 1: Purify Day 2: Buffer exchange and concentration Note : Both steps could be completed in one ...one (long) day. Equipment Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep (60% Iodixanol) QuickSeal... pooled for dialysis. Pool clean fractions. Concentration and buffer exchange: Stocks of Pluronic F68:...

... Protocols AAV Titration by qPCR Using SYBR Green Technology AAV Titration by qPCR...AAV Titration protocol. Protocols... Posts Mol Bio Protocols Introduction The AAV Titration protocol can be used to determine the number of...Size in bp of Addgene plasmid #59462 : 6208 bp Concentration of Addgene plasmid #59462 : 1.07 μg/μL Molecular...again but store at 4°C and use within 1 week. Keep track of the Ct value for each standard over time. They...over from the production process (DNase does not penetrate the virion). 5μL sample + 39μL H 2 O + 5μL 10x...sample (do not vortex) Incubate 30 minutes at 37°C Transfer to ice ** Critical: do NOT treat your plasmid ...

...including transformation, transduction, conjugation, and transfection. Transformation, transduction, and conjugation...created phages. Transfer of this DNA to another cell is referred to as transduction. Transferred DNA once inside...different methods of DNA insertion. Transformation Transformation is the uptake of genetic material from...fluoresce green. Comic by Maya Kostman.   Transduction Transduction occurs when foreign DNA or RNA is introduced...infected bacterium can either exist as transient extrachromosomal DNA, like a plasmid, or it can integrate...from a ‘helper’ phage prior to transduction. Scientists also use transduction to introduce foreign DNA into...donor can transfer genetic material to the recipient bacterium. The genetic material transferred is commonly...

...specific clone; lineage tracing is performed via barcode sequencing. Using the ClonTracer library, Bhang et ...interview with Novartis researcher, Carrie Bhang. The ClonTracer Library, deposited by Carrie Bhang, a research... exposure to the drug discovery process.” The ClonTracer library originally came about as Bhang and co-workers...questions that we could apply [it to].” Using ClonTracer to study drug resistance Although Bhang and co-workers...the publication, Bhang and coworkers used the ClonTracer library to determine whether or not cancer cell...the total population are easily be missed. The ClonTracer library allows a researcher to label individual...cells' genomes upon infection. When using the ClonTracer system, lentiviruses containing these barcodes...

...that the transformation and gene expression can be tracked. During a transient leaf transformation, the engineered... plant scientists do transient leaf transformations The development of transgenic plants is a common problem...method. Transfer your plant into the dark 16 hours before the transformation. We routinely transfer our plants...developing transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant...plant can take even longer, a transient plant leaf transformation process could save the plant biologist...development of transgenic hairy roots takes at least six months. Generating a transgenic plant takes even...reproducible. Prior to creating transgenic cultures or plants, our transient expression method enables us...

...opportunities to practice transferable skills in ways that will also enhance your science training and that will ... in a nonacademic direction, there are many “transferable” skills you have developed in addition to learning... concrete things you can do to develop those transferable skills while you are also learning to be an ...careers is that a team is almost always involved. Demonstrating your ability to work well with others can be...business” Attend events with local networking and trade organizations Another excellent perspective on ... transferable skills... science training...

.... This is why they are called transferable skills. Why are transferable skills so important? It is important... market, it is the transferable skills that create a full package and add an extra shine to your work.... be a more attractive candidate for that ideal job. How can you identify your transferable skills? PLoS...opportunities. Here are some strategies for identifying your transferable skills: Put yourself in situations...   What is a transferable skill? Time management. I needed it when balancing a handful of demanding courses...useful anywhere I work in the future; they are transferrable across most, if not all, industries and work...important to identify which transferable skills you possess because any potential employers, collaborators...