...Collections Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Additional Resources...Collections Background Tetracycline Off Tetracycline On Experimental Tips Tet Plasmids Background To advance the...induction with efficient tetracycline uptake. Search our collection for all tetracycline plasmids. Tet System...19 nucleotide tetracycline operator (tetO) sequence, and is recognized by the tetracycline repressor (tetR...tetR will bind to tetracycline and not to the TRE, permitting transcription. Tetracycline-dependent promoters...Bujard developed is known as tetracycline off: in the presence of tetracycline, expression from a tet-inducible... is reduced. To use tetracycline as a regulator of gene expression, a tetracycline-controlled transactivator...

...Monosynaptic Neuronal Tracing Viral Vector Packaging Service: Tools for Monosynaptic Neuronal Tracing These AAV encode...encoding tools for monosynpatic retrograde neuronal tracing, to be used with modified rabies virus. Viral...encode tools for rabies virus-based monosynaptic tracing. These AAV can be used with modified rabies virus...virus to perform retrograde neuronal tracing. Note : Addgene does not distribute rabies virus. AAV Vectors...Vectors for Monosynaptic Neuronal Tracing ID Name Description Serotype PI 52473 pAAV-synP-FLEX-splitTVA-...Introduction to Rabies Virus-based Monosynaptic Tracing The core idea of the system is to infect the cell... population of interest with a deletion-mutant tracing virus that lacks the ability to infect neighboring...

...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...

... Protocols RNA Extraction Without a Kit RNA Extraction Without a Kit...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...Figure 1: A diagram of the different steps in RNA extraction. Before Starting RNA is not as stable as DNA ... working with RNA, read this blog post on RNA extraction without a kit . Last Update: April 10, 2020 Equipment...additional steps to effectively lyse open the cells. Extract RNA from the homogenized sample(s).Transfer tissue...

...Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient Ultracentrifugation You may also...Follow this protocol using Iodixanol gradient ultracentrifugation to get pure adeno-associated virus (AAV) ... be completed in one (long) day. Equipment Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep...Data Figure 1: Iodixanol gradient before ultracentrifugation. The steps are clearly distinguishable thanks... 2: Left panel: Iodixanol gradient after ultracentrifugation. The arrow indicates the 60–40% interface...

...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...

...transsynaptic neuronal tracer into a monosynaptic (first order) neuronal tracer For RABV to crawl through...Retrograde neuronal tracing with a deletion-mutant rabies virus. Retrograde neuronal tracing with a deletion-mutant...neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability to efficiently enter cells...this transsynaptic spread is useful for neuronal tracing, it has two practical limitations. First, these...expression is a powerful development in neural tracing. Using these technologies in combination with molecular...neuronal populations. In one study, RABV monosynaptic tracing was used to examine how adult-generated neurons...the brain. Another study used RABV monosynaptic tracing to identify inputs to two major dopaminergic cell...

...expensive ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction. Researchers...your extractions- if you need really clean RNA, it’s important to make sure that when extracting, to only...contamination, take extra care when performing RNA extraction. RNA is inherently not as stable as DNA- it is... They will degrade in a -20°C freezer.   RNA extraction methods evolved into a simple protocol still ...without a kit. However, that isn’t the case for RNA extraction and purification. There is one simple method ... loaded onto cesium-chloride gradients and ultracentrifuged for up to 20 hours to separate the intact ...out to develop a protocol that skipped the ultracentrifugation altogether. Chomczynski and Sacchi showed...

... neuronal tracing experiments, brain tissue is injected with compounds that intracellularly spread away...further study. Traditional neuronal tracing methods Traditional tracing compounds either produce a colorimetric...anterograde tracer and injected at several hundred sites across the entire mouse brain. Traced neurons expressed...Background on neuronal tracing A key aspect to understanding the brain’s function is knowing its architecture...other distant brain areas. It is desirable for tracing compounds to outline neurons in the direction of...their signaling is likely to have). Anterograde tracing outlines neurons from their cell bodies to the ...the terminals of their axons; while retrograde tracing outlines neurons in the opposite direction, from ...

... tool that combines scRNA-seq and lineage tracking to track the dynamics of gene expression and clonal...combinatorial indexing strategy that simultaneously tracks clonal history and cell identity at the single-...with reprogramming outcomes (Biddy et al., 2018). Tracking lineage and cell identity at the single-cell level... of CellTag The Morris Lab developed CellTag to track the direct reprogramming of mouse embryonic fibroblasts...’s permissive to reprogramming.In this lineage tracing experiment, the Morris lab identified a novel gene...intestine, demonstrating its utility in long-term tracking in vivo (Guo et al., 2019). Key takeaways of CellTag...

... of XFPs in the original method, we used the Tetracycline trans-activator system to boost the expression...expression levels, and we named the system Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al.,... developing genetic tools for neuronal circuit tracing.     Marcus Leiwe is an assistant professor at ...about Brainbow Learn about using AAVs for neuronal tracing Visit the viral vectors blog topic page Additional...

...must occur by doing work outside of the lab. Extracurricular activities don't have to be ‘extra’ Today’s...in a myriad of professions is to engage in extracurricular activities (EA). EAs can: Introduce new perspectives...strengthen interests and skills. Although the word “extracurricular” may be taboo in the lab, multiple research..., TAKE ACTION! Taking action: Examples of extracurricular activities The table below lists potential...professions within and beyond academia. Extracurricular Activities Professional Skills Open source...

...contain plasmids deposited at Addgene from the abstract page. Check out that beautiful Addgene “Find Plasmids... Plasmids” badge in the image below. If an abstract on PubMed has the “Find Plasmids” badge, you’ll know... your PubMed account and are browsing article abstracts you should see the “Find Plasmids” box on papers...

... around or misread.  2. Track the container - It is just as important to track the location of the container...is to track the tubes themselves. Although we’ve developed our own computer system for tracking samples... a series of events is triggered. Each step is tracked through barcoded tubes that are scanned into our...Management System (LIMS), a database we use to keep track of and manage all of the samples that come through...be a nightmare if we didn’t have a simple way to track all of our 40,000 samples available for distribution.... This number ensures that every sample can be tracked easily at each step in the storage process. After...scanned to verify that it is the correct sample. Tracking samples - it's all about location, location, location...

...reagent storage and inventory, managing orders, tracking students and research projects, electronic lab...share protcols, sign-up for shared equipment, and track inventory and orders. Quartzy caters to life science...

.... Eyquem, Justin, et al. "Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection...

...Song, Shiping, et al. “Aptamer-based biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February...

...ATP (extracellular) ecAT3.10 extracellular ATP sensor (cell surface targeted) Imaging extracellular ATP...Yasushi Okada ATP (extracellular) GRAB_ATP1.0/ATP1.0mut biosensor for extracellular ATP (Constitutive ...Eckhard Boles pH Imaging intracellular pH in live cells (pHRed) Imaging Intracellular pH in Live Cells with...e.g. in vitro vs. in vivo , cytoplasmic vs. extracellular vs. organelle-localized). Browse or search the...lifetime-based biosensor for quantitative imaging of intracellular calcium. Nat Commun. 2021 Dec 9;12(1):7159. ... encoded fluorescent indicators for imaging intracellular potassium ion concentration. Commun Biol. 2019...Genetically encoded FRET sensors to monitor intracellular Zn(2+) homeostasis. Nat Methods. 2009 Oct;6(...

...discussing conversion tract length in drosophila. This 1998 study suggests mammalian cells' tract lengths seem ...recommends using the Epicentre QuickExtract solution to perform this step to extract genomic DNA from cells. ...reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01015-5 As for target selection,...reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01016-7 In the case where you have.... The Zhang lab uses around 50ul of the extraction solution, runs the protocol, then uses around 2-4ul...4ul in each 50ul PCR reaction. The gDNA extracted in this way is usually more concentrated than using other...pre-processing form of crRNA is expressed and a tracrRNA (the helper RNA that facilitate the processing...

... μl of DNA extraction solution. NOTE: Generally ~100,000 cells are used for DNA extraction, although a...loading this video, please try viewing on YouTube . Abstract The prokaryotic clustered regularly interspaced...following program: 65 °C for 6 min, 98 °C for 2 min to extract gDNA. Measure the DNA concentration. NOTE: While...and 6.2 recommend an efficient method for DNA extraction, any method for genomic DNA isolation may be ...flicking the PCR plate over a sink. Add 50 μl of DNA extraction solution per well and resuspend. Continue to ... cells to grow. Aspirate media. Add 50 μl DNA extraction solution per well, resuspend and transfer to ...to 96-well PCR plate. Continue to step 7.3 . Extract the gDNA from clones. Run sample in thermocycler: ...