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We narrowed to 881 results for: NSI

Showing: 351 - 400 of 881 results

  1. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...show a high density of cells, while contour lines that are spread apart show a low density of cells (Figure... a ‘gate’; it tells the machine that everything inside the gate is one population and everything outside...Heat maps are often used to indicate the relative density of cells. You can see both of these features in...

  2. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...vector into your target cells; you also have to consider how to keep it there. In fast-growing populations...needed to retain the vector as cells divide. In transient transfection, the vector remains episomal, meaning...about vector replication at all. In these cases, transient transfection may be sufficient for your needs ...involved in the production of viral vectors, we consider the viral vectors themselves to be a distinct ...

  3. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...that evolved in the last 10,000 years (which is considered recent). They have diverse traits with good genetic... and one is promiscuous and the gene that is responsible is understood. It’s possible to actually silence... to get a recombinant because this process is expensive. They are using a cool fluorescent indicator system...trying to replace embryo injection (hard to do, expensive, not every lab has the capability) with receptor...

  4. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...systems that are widely present in bacteria. They consist of a toxin which can affect a variety of cellular...division”). The encoded toxin-antitoxin system consists of CcdB (toxin) and CcdA (antitoxin) where CcdB...systems Because restriction modification systems consist of two enzymes that have opposing activities, it...the systems just described, the hok/sok systems consist of three components: Host killing (hok), a long-living...

  5. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...within Cas13a are essential for RNA cleavage, consistent with known roles for HEPN domains in other proteins...located at the 3’ end of the spacer sequence and consists of a single A, U, or C base pair. In bacteria,...bacterial cells will likely be limited by the propensity of Cas13a to bind and cleave RNA non-specifically...Finally (and perhaps most interesting), Cas13a’s propensity to cleave any RNAs after binding a user-defined...

  6. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...letters, while higher-level jobs are more likely to consider cover letters an important part of the application... content team, where writing is a core skill, considers cover letters to be very important. On the other...unofficial poll, hiring managers also said that they considered cover letters to be more important for higher-level...communication skills” try giving an example, like “I have extensive communication experience, having given more than...

  7. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...included tests done in cell lines known to be more sensitive to toxicity from RNA editing, such as mouse embryonic...CRISPR-driven precision into a genome, can be considered the fourth generation of CRISPR, one which drops...drops itself neatly into that gap. Consider, for instance, a disease like cystic fibrosis, which can be...that this system wasn’t dependent on the cell’s intrinsic machinery being already activated to work. (It...

  8. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...The ACE reporter is a bicistronic construct that consists of a mutated mCherry and a downstream constitutively...that results in the restoration of fluorescence. Transient reporter for editing enrichment (TREE) The Wang...Wang and Brafman lab developed a transient reporter for editing enrichment (TREE) to purify single base...Schwarz G, Nguyen T, Wang X, Brafman DA (2019) A transient reporter for editing enrichment (TREE) in human...

  9. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...Previously, we have described a number of practical considerations that should be taken into account when deciding...needs. Equally important, before cloning you must consider whether and how well your FP will function in ...fluorescent proteins in your experiments, please consider the following: How can one ensure that an FP ...compartments using CRISPR systems should strongly consider using moxFPs over standard FPs for the reasons...

  10. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...using microscopy techniques. FRET measurement sensitivity makes it suitable for studying interactions within...are difficult to overcome but they have to be considered when analysing data from your FRET experiments... The simplest and the most popular one is the sensitised emission method, where the donor is excited by...acceptor fluorescence, this method is also less sensitive to direct acceptor excitation artifacts and it...

  11. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...called a katana. Opto-katanin consists of two separate blue light-sensitive systems, VVDfast to control ...switch. You may be familiar with popular light-sensitive ion channels used to control activities like neuronal...in other contexts are sure to reveal even more insights. Super-duper lipid editor And if recruiting proteins...

  12. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ...mechanisms, or ascertain which integrins are responsible for specific ligand-binding functions, all key...microenvironment. The actin cytoskeleton exerts tensile force, transmitted through the integrin, where ...recently showed RGD-binding integrins demanded higher tensions before they would enable cells to spread, resulting...symmetrical circles. While elucidating the role of tension, however, the researchers realized they needed ...

  13. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...screening strategy. Restriction enzyme screening is sensitive to single base pair changes, but the edit must...should be designed solely for restriction enzyme sensitivity or resistivity, and thus be a marker for edited...you’re worried about off-target effects, you can consider using a version of Cas9 that has been specifically... check out or blog on optimizing this step, or consider enriching for transfected cells by flow sorting...

  14. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...system uses an scFv to amplify the fluorescence intensity of a tagged protein. SunTag has two components...scFv-GFP fusion bind the SunTag scaffold, the intensity of the fluorescent signal is boosted and enables...to the disulfide bond independent folding of intrinsically stable scFv. PLoS ONE 12:e0189964 . https://...which uses an scFv to amplify the fluorescence intensity of a tagged protein. Resources on Addgene.org...

  15. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...have, and we developed a great plan to start consistently knocking out high-quality videos for you all...two options: Film outside at a distance, or film inside with one person in a room. I wouldn’t be in there...live. After doing a bit of looking, I found an inexpensive HDMI transmitter that could transmit a signal... stand and mounted a monitor to it. It had an extension cord attached, with the option to mount a battery...

  16. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... clients focus on their experimental needs and consider a budget as a resource checklist. A budget is ...centrifuges, sequencers and analyzers, and any ultra sensitive equipment (RT-PCR, Microscopy, Fluorescence Scopes...awry when several factors aren’t taken into consideration. 1. Always focus on the science When negotiating...However extreme a solution may seem, you should consider it if it will help you achieve your research goals...

  17. A Control for All Seasons

    Type
    Blog Post
    ...the results across runs. If the results vary considerably, then the high degree of variability in the ...assays have their own unique controls that must be considered (Burry 2011). Perhaps the most critical are those...materials, lethality of knockdown, or other factors, consider using one of the alternative, albeit less ideal...signal remaining is autofluorescence and should be considered in the interpretation of results. Include the...

  18. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...first.  AAV-mediated gene expression can differ extensively depending on the experimental system used, so...Watakabe et al., 2015) (Figure 1). You’ll need to consider if such properties could help, or harm, your experimental...occur (Fig. 3) - for example, some neurons can transiently express TH - so you’ll need to check for this...injection volume.  There are a few factors to consider. First, the titer required is inversely related...

  19. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ... sequencing would be too expensive, then the qPCR method should be considered as an alternative. The qPCR...three-input GRSMs, we recommend using overlap extension PCR and Gibson assembly to piece together the ...circuits in general would benefit from a more comprehensive characterization of the recombinases at our ...

  20. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ... have engineered pADH1 variants that are less sensitive to ethanol, including the “middle” ADH1 promoter...systems. Binary systems are genetic tools that consist of two parts, both of which must be present for...results in chimeric GAL80-repressible and GAL80-insensitive activators of lexAop. This system is commonly...expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992 Jun...

  21. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...Application Recombination Frequency Special Considerations dsDNA Insertion or deletion of >20 nucleotides...CI. For recombineering purposes, a temperature sensitive version of the repressor gene, CI857, is used....express lambda red genes from a plasmid with a heat sensitive origin of replication.  Once the recombineering...expression of the int and xis genes which are responsible for excision of mini-λ from the host’s chromosome...

  22. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...encode a transposase enzyme.  Class I TEs are considered replicative since every time they jump they make...transcription start sites, CpG islands, and DNaseI hypersensitivity sites. Like Sleeping Beauty, piggyBac has ...transcription start sites, CpG islands, and DNaseI hypersensitivity sites. Tol2 is only active in vertebrates ...site preference and host range are important to consider when selecting a transposon for a mutagenesis ...

  23. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...life-saving research are being proposed.  It's our responsibility to join the public conversation and to tell...scientists – including global warming skeptics – insisted that more data and research funding are critical...towards starting these conversations; it is our responsibility to continue the dialogue. Erin Golden is currently...continue to worry) that most who participated will consider that marching in and of itself will be enough....

  24. Typing CRISPR Systems

    Type
    Blog Post
    ...and understanding of CRISPR, and perhaps provide insights and ideas for your own research! Classification...with BioRender.com.   Type III Type III is considered the most complex of the CRISPR types and is thought...et al., 2021). IV-A and IV-B lack what you may consider to be an integral component of CRISPR — a nuclease...2021). Evolution of Type IV CRISPR-Cas Systems: Insights from CRISPR Loci in Integrative Conjugative Elements...

  25. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...wherein multi-part assembly can occur. It is quite extensive with a supplemental add-on kit and is compatible...William M., et al. “A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces Cerevisiae....pone.0197185. Chamness, James C., et al. “An Extensible Vector Toolkit and Parts Library for Advanced...2024.02.16.580396. Martella, Andrea, et al. “EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid...

  26. Addgene’s New Way to Help You Share Science

    Type
    Blog Post
    ... with “A Better Way to Share Science” for: The inside scoop from scientists who have deposited plasmids...latest technologies and breakthrough research. Insights about how to navigate through all stages of your...and Facebook. We’ll track down the story, get the inside scoop, and keep you informed.  ...

  27. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    ...Listen to this episode for insights into the process of getting an MD PhD, the inside scoop on developing new... MD PhD we've had on the podcast, has worked extensively with CRISPR, and recently became a member of ...

  28. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...and on-targeting likely more efficient.  These insights are hard to appreciate without computational support...tools The goal of this post is to provide some insight into available CRISPR software tools, what problems... software tools that apply some approximate or sensible rules to determine the value of one sgRNA design...functions are not entirely necessary but should be considered extremely useful, depending on design goals. ...

  29. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

    Type
    Blog Post
    ...transformation into Sample 6 and Dr. Koeris' own transition from graduate student to entrepreneur, listen...founding a startup company? 32:20 - 34:37 : Whose responsibility is it to take on the risk of developing products...37-36:53 : Was It difficult for Dr. Koeris to transition from developing therapeutics to developing a ...

  30. A Scientist's Guide to Ebola

    Type
    Blog Post
    ...week within two months. That said, Ebola is not considered a “terribly infectious disease” – infection occurs...via body fluids not through the air and that’s considered unlikely to change – but the illness has been... critical for viral attachment and fusion. Once inside a host cell, new evidence in PLoS Pathogens shows...

  31. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    ...quoted as saying when the news first broke. "Considering that many human diseases are caused by genetic...presented the first detailed picture of Cas9's three-dimensional shape via x-ray crystallography, an advance they... and mainstream the news on CRISPR has gotten, consider these headlines in The New York Times and Forbes...

  32. Download Addgene’s Science Career Guide, Plus 8 Career Tips from Addgenies

    Type
    Blog Post
    ...compiled our top career blog posts into one comprehensive, downloadable guide: the Addgene Science Career...you’d like to see discussed on the Addgene blog, consider writing a guest post. Additional resources on... Find job listings at Addgene Get more career insight from Addgenies ...

  33. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    ...right! He began to study the A. victoria gene responsible for encoding GFP, and, in 1992, he reported its...hypothesized that GFP-like proteins could be responsible and began studying these marine organisms to ... fell into place, as they, taken alongside the insights provided by the studies in Tsien lab, covered ...

  34. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...number of building blocks with just two or three inexpensive enzymes. Now, a new toolkit - aptly named GreenGate...endonuclease of choice, BsaI, which can be very sensitive to temperature fluctuations. He says their lab...problem back to the endonuclease. “It’s just a sensitive protein,” he says. “There is nothing you can do...

  35. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...the cell density and viability: Using a 5 mL pipette, transfer 0.5 mL of HEK293 cell suspension into a ...microcentrifuge tube. Vortex the cell suspension. Transfer 10 µL of cell suspension into the microfuge tube containing...Protocol Introduction Transfections allow for transient expression of a gene of interest in cell culture...culture. This protocol describes how to transfect suspension HEK293 cells with recombinant antibody plasmids...support reproducibility in science. General Considerations For antibody-expressing plasmids containing...antigen (EBNA). Safety Warnings HEK293 cells are considered biosafety level 2. Please ensure that you are...transfecting, seed a 108 mL culture of HEK293 cells at a density of 0.9 x 10 6 cells/mL in a 500 mL vented flask.Incubate...

  36. Scientist Networking: What is an Informational Interview?

    Type
    Blog Post
    ... from other scientists doing interesting work. Consider this to be like any other research project. You...develop relationships, but if it is not possible consider a phone or video call. If you do decide to use...personal questions can elicit the most valuable insights you will get from the interaction. Most people...

  38. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...organism has already been made, and it would be expensive and time-consuming to repeat the process with ...spontaneous conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White...level necessary to observe a signal. Red signal intensity increases with GFP concentration and saturates...

  39. 2023 and 2024: Reflections and Looking Ahead

    Type
    Blog Post
    ...partners, with the launch of antibodies in China and expansion of viral services available in South Korea.  ...addition, we released the MORF collection, a comprehensive transcription factor library, in lentiviral ..., Korea, Japan, India, and Brazil. Through an expansion of our export license application service, we’...

  40. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...a new and versatile binary Ti vector for both transient and stable gene expression applications in plants... (GUS) and GFP reporters, as well as multiple transient and transgenic systems, Sahoo et al. showed that... a novel versatile gene expression vector for transient assays as well as stable expression of foreign...

  41. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ..., yellow, cyan, blue, and orange spectra. A comprehensive list of cell markers all tagged with mEmerald...multicolor labeling and FRET. Specialty constructs for biosensing. Over 300 plasmid backbones spanning a rainbow...color variants. The mEmerald plasmids offer a comprehensive list of readily available targets since the ...

  42. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    ...fluorescent proteins to observe what’s happening inside cells for at least a couple of decades. Green is... PA FPs with light of specific wavelength and intensity allows for optical labeling and tracking of proteins...capable of revealing, for example, how muscles work inside a living animal. For those making use of these ...

  43. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ... these techniques and is a multi-step process consisting of digestion, purification, ligation, and transformation..., TOPO cloning, one step cloning, and overlap extension PCR. Although each type of cloning has its advantages...combine the products and digest with methylation sensitive restriction enzyme DpnI to remove the templates...

  44. Embracing Serendipity: A Crucial Element in the PhD Journey

    Type
    Blog Post
    ...expanding your knowledge base extensively, you open doors to fresh insights and broaden your understanding...Making the Leap to Industry." He successfully transitioned to industry after his PhD and a couple of postdocs...

  45. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    ...put on typical middle age weight and “exercise” consisted of taking walks in the mall. I took action, joined...patients. However, the R206H mutation makes ACVR1 responsive to members of the ubiquitous activin ligand family... to raising funds and awareness of FOP. Please consider running with us or making a donation via the link...

  46. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    ...for the user. This approach serves to maintain consistency with published screen interpretations and allows...curated CRISPR screen data are also available via comprehensive pre-compiled downloads and via a full-featured...are fully open access and available without any licensing restrictions.    As CRISPR technology continues...

  47. The Stingy Scientist: How the Baby Gel Box Was Born

    Type
    Blog Post
    ...labs. Disposable plastic materials are fairly inexpensive in this country and we are quick to throw out... gel on a large glass slide using only surface tension.  It's a bit tricky to pour the right amount so...plastic and solder this thing together.  One of the downsides—there was no cover so you had to be careful not...

  48. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...how Protein A and Protein B interact. You read extensive research on the two proteins and come up with ...antibody against Protein B. The antibodies are both extensively published, knockout validated, and work in your...with human antibody sequences.  Additional considerations Although isotype conversion is routinely used...

  49. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ...markers and replication origins, but there is extensive heterogeneity in the feature sequences due to ...control elements in commercial plasmids were inconsistent and sometimes clearly wrong. In an effort to...reliable feature annotations. Even with these extensive efforts, such an algorithm has limitations. For...

  50. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...a relatively pH-insensitive RFP variant, DsRed.T3, that’s connected to a pH-sensitive GFP variant, super...lysosome, and degradation of autophagic substrates inside the lysosome. Historically, methods for studying...
Showing: 351 - 400 of 881 results