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We narrowed to 435 results for: Trac

Showing: 1 - 30 of 435 results

  1. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...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...

  2. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ...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...

  3. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...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...

  4. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ... 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 ...

  5. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ... 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...

  6. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ... 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...

  7. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    Published
    Jan. 21, 2014, 3:08 p.m.
    ...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...

  8. Activate Addgene's "Find Plasmids" Badge on Pubmed Abstracts

    Type
    Blog Post
    Published
    May 12, 2017, 12:58 p.m.
    ...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...

  9. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ... 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...

  13. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ... milk going to help me extract DNA from agarose gel?!”   The DNA gel extraction with glass milk method... a similar workflow as a column-based plasmid extraction kit. First, you lyse the bacteria and denature...contamination and you can use a phenol chloroform extraction to remove remaining proteins and RNase. Finally...ready for use! Plasmid purification and DNA gel extraction with glass syringe filters Starting material:...fragments The plasmid purification and DNA gel extraction method described in Kim and Morrison is nearly...protocol and guanidine isothiocyanate for the gel extraction protocol) disrupts the DNA’s hydrogen bonds with... more flexible than commercial kits.  DNA gel extraction with glass milk Starting material: agarose gel...

  14. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    Published
    April 27, 2023, 1:15 p.m.
    ...information from a region (anterograde tracer, Fig. 1B). Tracers are typically delivered to a particular... either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...making them useful tracers, but finding a virus that works well for retrograde tracing can be difficult....brain works. There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information...information to a region (retrograde tracer, Fig. 1A), and another to visualize neurons that receive information...from the axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses...transport and can therefore perform retrograde tracing, but often at the cost of toxicity, low expression...

  15. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...days. The crRNA and tracrRNA are annealed on a PCR block and the annealed cr:tracrRNA is then mixed with...Anneal crRNA and tracrRNA using a PCR block. Add Cas9 protein to the annealed cr:tracrRNA and incubate at...Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP) delivery Epitope...started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments (in collaboration with ...-tagging experiments. Cas9 protein, annealed cr:tracrRNA, and single-stranded donor DNA. The success of...have recently demonstrated that the two-part cr:tracrRNA system is much more potent than the in vitro-...Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient, it is also straightforward...

  16. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ... challenging to track intracellularly, but aptamers permit the study of intracellular RNA dynamics. Aptamers...its ability to track RNA dynamics in a model of Fragile-X tremor/ataxia syndrome (Strack et al., 2013)....target cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex... molecule. Fluorescent RNA aptamers are used to track cellular processes by transforming cells with plasmids...then, Spinach2 has been used to investigate intracellular viral DNA dynamics in neuronal cells (Nilaratanakul...is better able to maintain its fluorescence intracellularly due to a lower dependency on magnesium and ...bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology 9.10 (2014): 2412-2420....

  17. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    Published
    Oct. 2, 2025, 1:15 p.m.
    ...Fusion Vector at Addgene! Following extracellular RNA editors Tracking factors that utilize exosomal transport...activity is compatible with both intra- and extracellular environments, making it effective for analyzing... overexpression to CRISPR knock-in, letting you track endogenous protein activity and measure responses... hypothesized that if ADAR1 were carried by extracellular vesicles (EVs), edited transcripts could endow...highlights HiBiT tagging as a sensitive reporter for tracking proteomic cargo flow via EVs. Find ADAR-HiBiT ... endocytic trafficking in real time through extracellular-specific and full lytic measurements. Find CFTR-HiBiT...CFTR-HiBiT Fusion Vector at Addgene! Real-time tracking of receptor internalization Work conducted internally...

  18. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    Published
    Nov. 3, 2016, 2:30 p.m.
    ...video abstracts identified. The journal Cell had the largest average view count per video abstract at 2,947...antibiotic prescriptions for respiratory tract infections. The video abstracts provide a brief introduction to ...science communication through video     Video abstracts about research, especially published research,... views) rounded out the top four average video abstract view counts.” For researchers looking to garner...garner more attention for their work, a video abstract may be a useful tool for turning video views into ...there was a growing number of online research abstracts. With journals in medicine, physics, math, and...), I created a new platform for sharing video abstracts about research: www.WeShareScience.org. WeShareScience...

  19. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...of common genetically encoded neuronal tracers Neuronal tracers such as epitope tags and fluorescent proteins...molecules in your favorite neuronal population and track connections to that population. Limitations of ... used for protein localization, isolation, and tracking experiments (Rizzo, 2009). With many different...Spaghetti Monster - A new and improved neuronal tracer With the limitations of epitope tags and fluorescent...fluorescent proteins, it has been crucial to develop new tracers that enable accurate labeling of neurons. As discussed...expressed proteins in particular are difficult to track with these classic probes. To overcome the limitations...Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins such as GFP have been used...

  20. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ...called Simultaneous Tracking of Recombinase Activation and Clonal Kinetics (STRACK) to study stem cell...several viral tools can perform neuronal tracing, anterograde tracing poses challenges, from non-selective...While many barcoding libraries are available for tracking cell lineage, different applications require careful...pre-existing heterogeneity of hematopoietic stem cells and track clonal dynamics during differentiation after they...labeling, making it a powerful tool for circuit tracing and other neuroscience applications (Figure 4).... single synapses, a key attribute for neuronal tracing. Labeling was nontoxic and activity-dependent, ...itself could be expressed Cre-dependently to allow tracing exclusively from genetically determined cells. ...

  21. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...available for tracking certain cell types. Another useful tool for marking and tracking cells is the the...fluorescently labelled antibodies made it possible to track cells from one organ to another. Thus several labs...lymphoid organs did in the past. How do researchers track fluorescently labeled cells? 1. Epifluorescence ...fluorescent proteins gave researchers the ability to track immune cells within organs and visualize how cells... to the immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting...after antigen challenge, scientists were able to track where GFP labelled B cells were in mounted slices...organs. These GFP labelled cells could also be tracked by cell sorter giving immunologists the ability...

  22. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    .... Developmental biology insights using lineage tracing in zebrafish To highlight some of the features ...zebrafish useful, I’ll outline two recent lineage tracing approaches that led to important insights in developmental...developmental biology. Microscopy-based lineage tracing Over a period of 24 hours, the externally fertilized...cells in the developing zebrafish embryo have been tracked using microscopy and advanced computational approaches...Keller et al., 2008). This microscopy-based lineage tracing defined a high-resolution map of early embryogenesis... mutagenesis screens. Microscopy-based lineage tracing can help illuminate how signaling pathways—such...development. Single cell sequencing-based lineage tracing Recently, gene expression in all cells of the developing...

  23. Build Your CRISPR Vocabulary

    Type
    Blog Post
    Published
    July 2, 2024, 1:15 p.m.
    ...wanted poster that Cas9 uses to track down its suspect—the target DNA. tracrRNA: A “trans-activating CRISPR... of CRISPR. The bacterial chromosome encodes a tracrRNA (in some systems including Cas9), Cas proteins...here). The repeat sequence base pairs with the tracrRNA, and the spacer sequence is used to target complementary...laboratory settings, an sgRNA includes the crRNA and tracrRNA sequences in a “single-guide RNA” that performs... crRNA, forming the repeat:antirepeat duplex. tracrRNAs also contain multiple stem loops that are recognized...proteins, like LbCas12a or AsCas12a, do not require a tracrRNA. sgRNA: “Single-guide RNAs” were invented by synthetic...synthetic biologists. An sgRNA combines a crRNA and a tracrRNA into a single strand that folds back on itself...

  24. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...its native context, the tetracycline repressor (TetR) can bind to the tetracycline operator sequences (TetO...preventing transcription. In the presence of tetracycline (Tet), TetR preferentially binds Tet over the...repressible system, Gossen and Bujard created the tetracycline-controlled transactivator (tTA). They fused ...TRE)), allowing transcription to proceed. When tetracycline or one of its derivatives is added, it binds...information on Addgene Tet plasmids, see our Tetracycline resource page. Cumate Switch Massie et. al developed...permit exquisite control of gene expression and tracing of gene expression across development. One common...control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S...

  25. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...a look at some popular self-labeling tools: Tetracysteine: The first reported chemical labeling tag, from...Commercial ligands available Ease of use Tetracysteine (Cys4) 15 Biarsenical dyes (FlAsH, ReAsH... HaloTag for their "ticker tape" method for intracellular recordings, switching between different colored...time points.   Figure 3: Single-molecule tracking of actin filament assembly-regulating proteins...appearance at the filament tip from (a). c) Intensity traces at the tip of the filament from (a-b), showing ...length (lower). d) Intensity and filament length traces for a different event where CP appears briefly ...labeling specificity than short peptide tags like tetracysteine (Jing & Cornish, 2011). HaloTag and SNAP-tag...

  26. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...difference. Traces The Traces tab shows the edited and control, non-edited Sanger traces in the region...as determined by comparing the edited trace to the control trace. In the ICE algorithm, potential editing...and Sanger sequenced. ICE compares these sequence traces to give a detailed analysis of CRISPR editing. ...three tabs - Contributions, Indel Distribution and Traces - provides particular details about the indel profile...the reference sequence derived from the control trace file. On the plot, the green (edited sample) and...calls from the .ab1 file are also shown above each trace. The horizontal black underlined region represents...

  27. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ... epitopes of interest that are found in the extracellular space. The targeted epitopes can be motifs within... T cell. Created using biorender.com.   Intracellular labeling Oftentimes, a marker of interest is... cannot penetrate the cell membrane; hence, intracellular labeling with antibodies requires chemically...analysis to be shifted to a later day. Note that intracellular labeling must be performed only after viability...staining and surface labeling are already done. Intracellular labeling can be subdivided into two categories...procedure in that case.     Figure 3: For intracellular labeling cells need to be fixed and permeabilized...cell proliferation and cell viability. Unlike intracellular labeling, dye staining can be done on live cells...

  28. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ... beliefs. I asked participants to read short abstracts comparing the mortality rates between two controversial...peer-reviewed study. The second group read the same abstract but with the names reversed, meaning that the ...reversed. The third group read a version of the abstract where the real medical conditions had been swapped...asked to evaluate the quality of the study those abstracts represented. The point of this study was to see...see if their evaluations of the abstracts were driven by the quality of the study design or by their preconceived... significantly more favourable ratings to the abstracts that affirmed their personal beliefs even though...identical. Likewise, when participants read an abstract for a study with findings that challenged their...

  29. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...conversion tract length in Drosophila. Elliot et al. (1998) suggests mammalian cells' tract lengths seem...recommends using the Epicentre QuickExtract solution to perform this step to extract genomic DNA from cells. .... The Zhang lab uses around 50 µl of the extraction solution, runs the protocol, then uses around 2–4 ...4 µl in each 50 µl PCR reaction. The gDNA extracted in this way is usually more concentrated than using...pre-processing form of crRNA is expressed and a tracrRNA (the helper RNA that facilitate the processing... then when expressed in cell and with Cas9 and tracrRNA, it will be processed (i.e., cleaved) into a shorter...form of crRNA is fused chimerically to a mature tracrRNA), the spacer used is only 20 bp. Please refer ...

  30. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ... and recent advances have allowed scientists to track multiple genomic loci over time using all the colors...CRISPRainbow. Figure 1: CRISPRainbow was used to track the localization of 6 chromosome-specific loci in...labeled dCas9 to a given genomic locus, you can track the localization and movement of this locus in live...depositor Stanley Qi and used in Chen et al. to track telomere dynamics throughout the cell cycle. To ...distances from 2-75 Mbp. Labeling multiple, intrachromosomal loci Building on the work of Chen et al., ...turned their attention to mapping pairs of intrachromosomal loci. The technique successfully resolved ... this work represents the first mapping of intrachromosomal loci, a major benchmark in characterizing ...
Showing: 1 - 30 of 435 results