...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...

... 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...

...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...

...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...

... 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....

...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...

...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...

...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...

.... 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...