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  1. Light Sheet Fluorescence Microscopy

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    Blog Post
    ... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...multiplying couple-charged device (EMCCD) or complementary metal-oxide semiconductor (CMOS) sensors, it...labeling in whole tissue (15).  CLARITY is also compatible with LSFM, a method termed COLM (clarity-optimized...spatiotemporal control over labeling methods as compared to transgenic animals (17).  These viral plasmids...
  2. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    Blog Post
    ...For example, Drosophila lines with compound autosomes are completely infertile when mated to wild fruit...with appropriate measures. Barriers should be a component of all gene drive confinement strategies, but ...: I. Cage Studies of Chromosome Replacement by Compound Autosomes in Drosophila melanogaster. Genetics...
  3. Transferable Skills Guide: Identifying Your Transferable Skills

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    Blog Post
    ...this situation: You’ve spent hours scouring a company’s LinkedIn pages, given yourself the bathroom mirror...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...the candidate has what it takes to thrive at the company and add value, not just “do the job”. When they...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...
  4. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ... an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused to an RNA adenosine...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...
  5. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

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    Blog Post
    ...orders were affected by the COVID-19 pandemic, we compared the number of plasmid orders and new COVID-19 ...the likelihood in our Poisson process into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential...effects fairly well. However, we observed one complication in the China data – there is a period of 1 month... lockdown in China, then the observed data is compatible with an infinity-fold reduction in activity due...
  6. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...real time. When the PCR is complete, the results are analyzed by comparing the threshold fluorescence ...amplification reaction to take place.  When the PCR is complete, a droplet reader extracts the droplets from the...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...
  7. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...them to near homogeneity using a 6XHis tag, and compared their specific activities by measuring NADH production...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...
  8. AAVs CREATed for Gene Delivery to the CNS and PNS

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    Blog Post
    ...gross comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids....PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived from. They also...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...
  9. Lentiviral Vector Uses and Overview

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    Blog Post
    ...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...system, the possibility of creating a replication-competent virus via recombination between the transfer and...recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition...Lentiviral barcoding libraries have been used for complex applications like lineage tracing in heterogeneous...
  10. Plasmids 101: Colony PCR

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    Blog Post
    ...restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money... few ways you can do this. If you are going to complete your colony PCR analysis in the same day, you ... product size on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel... larger sized products for positive clones (+) compared to negative clones. Finally, orientation-specific...
  11. Tips from the Repository Trenches: Using Barcodes to Track Samples

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    Blog Post
    ...is ideal for tracking large collections. Many companies, such as Thermo Fisher and Micronic, provide 2D... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...specific barcodes etched or printed onto them by companies such as Tubewriter. Any lab can benefit from using...tubes themselves. Although we’ve developed our own computer system for tracking samples, it can be done easily...
  12. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...survive as a researcher because of the tough competition for academic positions and poor grant situation...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...
  13. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

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    Blog Post
    ...on OWW, including those for preparing your own competent cells. If you’re familiar with the process of ...service free of charge to scientists by allowing companies to pay publish their catalogs on the site. Like...of checking off each step as you painstakingly complete your experiment – one small victory in a long ...manage scientific data: Electronic lab notebooks compared with Google Docs and wikis - by Rory Macneil on...
  14. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn...variants that reached the CNS following IV injection. Compared to AAV9, PHP.B has a >40-fold increase in CNS ...rat CNS: AAV PHP. EB vector dose–response and comparison to AAV PHP. B." Gene therapy 25.5 (2018): 392...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...
  15. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...are needed to deliver all the protein and RNA components, and several sgRNAs must be screened to identify...eBook Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...
  16. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...in order to be expressed stably in neurons. As compared to epitopes tags, FPs are intrinsically fluorescent...fluorescent proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags. Indeed,... structure resembles images of the deity (see comparison image above). The church of the Flying Spaghetti...
  17. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...
  18. Plasmids 101: Gateway Cloning

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    Blog Post
    ...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...
  19. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    ...target provided that (1) the sequence is unique compared to the rest of the genome and (2) the sequence...for the canonical S. pyogenes PAM sequence NGG compared to wild-type SpCas9, which also displays some ...CjCas9) is even smaller than SaCas9 and is also compatible with AAV delivery. NmCas9, another small ortholog...(2018) Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 ....
  20. Hot Plasmids - August 2020

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    Blog Post
    ...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate...
Showing: 501 - 520 of 928 results