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We narrowed to 881 results for: NSI

Showing: 401 - 450 of 881 results

  1. 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...

  2. 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...

  3. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    ...at our graphics and design editors. How about condensing a week’s worth of the world’s scientific discoveries...like word choice, punctuation, and stylistic consistency. If you’ve ever doubted the importance of copy...in the science communications world, you might consider an internship or a short program to build skills...

  4. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    ...gene expression in a target cell. Practical considerations If you have high quality AAV, you can address...required for the transduced cell to become light responsive than fluorescent proteins required to get a good...interior, causing less particles to be released into suspension and lowering the number of viral particles that...

  5. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...space where BosLab exists today. Back then, it consisted of “wooden slab benches” set on top of filing ...companies and BosLab has grown into an organization consisting of over twenty members and volunteers. Transforming...biologist with a Ph.D. from UNC-Chapel Hill and has extensive experience targeting RNA with small molecules....

  7. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ...caspases are activated last in the cascade and are responsible for the degradation of over 600 cellular components...detected fluorescence within 2 to 5 hours, a time consistent with previous methods. In zebrafish, the Shu ...developing forebrain and retina. This pattern was consistent with TUNEL staining, a popular method to observe...

  8. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...using CRISPR/Cas9. They are targeting the genes responsible for both early onset and late onset AD. In this...These papers suggest it might work well enough to consider as a therapeutic.” There are many fundamental ...handle on how cleavage sites are repaired.” Those insights will be critical for scientists’ ability to influence...

  9. The MTA at Addgene

    Type
    Blog Post
    ... institutes, the Tech Transfer department is responsible for managing research use agreements like MTAs...beginning makes the sharing process smoother, more consistent, and more pleasant for everyone, which in turn...including MTAs, is perfect, but it does provide consistency and transparency in an otherwise messy process...

  10. Hot Plasmids - January 2023

    Type
    Blog Post
    ...applications such as super resolution, electron, and expansion microscopy. hfYFP is not only brilliantly bright...than most FPs. The group also tested hfYFP in expansion microscopy as well as correlative light and electron...stable FP for your next microscopy experiment, consider hfYFP and LSSmGFP.     Fig. 4: hfYFP retains...

  12. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    ...found, to providing reagents in an accessible and consistent way so other researchers can use them, to making... can and do play this role, and we take our responsibility to push for clear communication of science ...broader role as Vice President. Her current responsibilities encompass overall leadership of the Cell Press...

  13. How to Lead a Great Meeting

    Type
    Blog Post
    ...are there. Make the most of everyone’s time by considering the level of meeting you need. Project Meetings...meetings tend to fill the shape of their container. Consider whether you actually need a full hour to achieve...FInd Tips on Picking a Research Project Read Other Inside Addgene Posts Resources on the Addgene Website...

  14. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ... perhaps surprising, yet important, factors to consider when seeking your dream lab. The principal investigator's...want a non-academic career path?  If you are considering this, make sure to choose an advisor that supports...where the topic of your research can make your transition easier – choose a lab that works on problems ...

  15. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ... Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than...Red flame plasmid pDawn contains the blue-light sensing protein YFI. When light is absent, YFI phosphorylates...? Here’s a list of some important variables to consider when choosing an inducible promoter. Experimental...

  16. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...this solution wasn’t practical due to the low intensity of luminescent proteins. To make luminescent imaging...luciferase has a low quantum yield, and when transiently expressed in cells, generates much less signal...intravenously with coelenterazine, with increased sensitivity and faster imaging compared to previous publications...

  17. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ... lower background levels and improved signal sensitivity. Another advantage: luciferase reporters aren...Not required Signal strength Lower Higher Sensitivity Higher Lower Background Lower Higher Photobleaching...change in conformation of a single protein. Considerations for luciferase assays Luciferases can only ...

  18. Clear and Accessible Writing

    Type
    Blog Post
    ...macromolecule density is equal to the buoyant force of the solution. Contaminants of lower densities are captured...did! From Addgene's very beginning, we had to consider digital accessibility for our website and online...

  19. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ... mammals, which is thought to be due to their propensity to integrate into proto-oncogenes and disrupt...alternative virus, or other strategies may need to be considered if size constraints are an issue. Adenovirus...Adenovirus is an effective option if you need high, transient expression of a genetic cargo or have a very large...

  20. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...repair template (sister chromatid or homologue) or extensive DNA synthesis. NHEJ also finishes repair of most...streamlined version of this pathway and does not consider the missing or damaged nucleotides that are common...Biochemistry and Biophysics and the Lineberger Comprehensive Cancer Center at UNC Chapel Hill.   References...

  21. How to Make Friends and Meet People at a Scientific Conference

    Type
    Blog Post
    ...you are spending on a conference? One thing to consider is the size of the conference. Large conferences...favor small conferences where the attendees stay onsite and eat together because scientists have time to...– even students and postdocs. There are many inexpensive/free places to order cards. There are a few good...

  22. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    ...publication in a scientific journal requires several considerations. First, it is important to establish the type...journal. An increasing number of funding bodies insist that the research data resulting from their funding...is a Developmental Editor for F1000Research, responsible for assessing the scientific content of manuscripts...

  23. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...red-shifted Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent... Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin Ace2N and mScarlet. Ace2N...existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins. Find the Ace-mScarlet voltage...

  24. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...sequencing and qPCR methods are too costly and time intensive to process a large number of microbiome samples...bumble bee and honey bee microbiome. The toolkit consists of modular parts that are assembled with Golden...microbial population dynamics in the gut can provide insight into how the microbiome responds to infection, ...

  25. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ...antibody, this approach can greatly increase the sensitivity of an assay by increasing the number of signaling...place over multiple days. Increased run time is considered one of the major drawbacks of the indirect method...interest - the secondary antibody needs careful consideration before selection. If all the primary antibodies...

  26. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...advantage of polyclonal antibodies is that they are inexpensive. Their short production time and the bulk purification...labs. Polyclonal antibodies also have a high sensitivity to the target protein due to their ability to...or fixation (IHC). Because of their increased sensitivity, polyclonal antibodies are particularly useful...

  27. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...might use it in the lab. What is a Fab Fragment? Consider the classic IgG antibody (Figure 1): it is composed...new components in your antibody tool box. As you consider which tools to use, be sure to think about what... other components of your experiment. These considerations will help you narrow down which antibody format...

  28. The time and cost required to make a plasmid

    Type
    Blog Post
    ...organized lab saves time and money across the board. Consider alternative cloning strategies There are many ...methods, like Gibson assembly, are typically more expensive, so you’ll have to find a balance between time...either don’t scale or have limited scaling. The downside of high-throughput work is that you give up a ...

  29. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ... dsDNA. It cleaves ssDNA with high fidelity - sensitive to even a single mismatch in the target sequence...This makes it particularly useful in generating transient edits of signaling molecules (RNA) as opposed ...for targeting mRNA and useful as a mechanism of transient knockdown of gene expression. This system could...

  30. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    ...using words for their field or technology in a consistent way. Just creating a lexicon was not enough. ... using the same words with the same meanings consistently, meetings will be shorter and more productive...during your postdoc that you enjoy and have a propensity for cross team communication, that’s an important...

  31. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ...negative regulation by using a promoter that is responsive to a repressor protein, the increase in expression... of repressor protein also increases thereby intensifying repression of the gene of interest and keeping...the increase in plasmid copy number during the transition to stationary phase, Segall-Shapiro et al. placed...

  32. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...that consistently receives around 200 new DNA samples every week. Here we will provide an inside look ...terminators. Detailed instructions on other factors to consider when designing your primers can be found here....

  33. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...site-specific recombination events. The system consists of two components derived from the P1 bacteriophage...cyclization recombinase”), is a 38 kDa protein responsible for intra- and inter-molecular recombination ...sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats separated...

  34. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... short-term protein expression observed using transient transfection approaches, generating cell lines...experiments in a stable cell line, as opposed to transiently-transfected cells, increases reproducibility,...eliminates the variation associated with repeated transient transfection. Some lentiviral vectors deliver ... in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health of the target cell...pipetting or inverting the tube. Aliquot 1 mL of cell suspension (i.e., 50,000 cells) into each well of the 6-...the virus will have higher levels of expression. Consider expanding populations transduced with a variety...expressors may take over the culture. To overcome this, consider generating monoclonal lines from the early polyclonal...

  35. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ... publication, Mary Dunlop’s lab generated a photosensitive Cre, Opto-Cre-Vvd, by splitting and joining.... Opto-Cre-Vvd is precise, demonstrating low sensitivity to ambient light, and versatile as it can be ...be activated by both low and high-intensity blue light. Unlike other split-Cre systems which can take up...

  36. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...Performance When targeting single genes, SAM consistently shows the highest levels of gene activation ...which are relatively large and are not expressed consistently throughout cells. VPR Description VPR fuses ... dCas9 to activate transcription. This complex consists of the VP64 activator used in other CRISPR activation...

  37. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...position, I just had a phone interview. I also transitioned between different lab groups without having ...that have lots of general questions about job responsibilities, performance evaluations, professional development...difficulties. Have your presentation on a flash drive and consider bringing your own laptop as a backup, if the company...

  39. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...science and collaboration. The field already has comprehensive reference pages and curated databases for scientists... (CeNGEN) The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron...Scientists monitor the effects of RNAi using four-dimensional contrast interference (DIC) microscopy and computer...

  40. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ... this type of ELISA is extremely specific and sensitive to the target protein due to the use of multiple...concentrations of your protein of interest. The optical density (OD) - the measure of light absorbed by a solution...any experimental wells in that range of optical densities. If your standard curve does not have a linear...

  41. A Look at Addgene's QC Process

    Type
    Blog Post
    ...significant plasmid elements. Which differences are considered a QC issue is reviewed yearly at a best practices...populations. All of these topics require additional considerations to resolve. Sequencing and assembly issues ...assembly in Geneious can still provide additional insights to resolve the assembly and determine if there...

  42. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...article we will discuss general viral safety considerations and best practices for handling lab-grown viruses...replicate and continue the infection cycle once inside the host. While the virus can still enter (and ...into the host’s DNA. These viruses also have a propensity to integrate into proto-oncogenes, which can ...

  43. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...fluorescent in-situ hybridization, methylation-sensitive restriction enzyme digestion, DNA adenine methyltransferase...living cells. Essentially this reporter construct consists of one substrate (the histone-derived peptide)... of these sites was controlled, the reporter consistently showed significantly different FRET levels between...

  44. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...requires a camelid animal facility which can be expensive to gain access to routinely, and it can be especially...the mRNA molecule, forming a ribosome complex consisting of the translated protein (sybody), ribosome,...constructed, they are used to produce a phage library consisting of phages that display the candidate sybodies...

  46. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    ...poster – now it’s time to practice your delivery. Consider your audience: Who will be attending the session... for Cancer Research Annual Meeting, you might consider including more background on how your virus of...practice: Rehearse your presentation with people inside and outside your field. Your colleagues, lab mates...

  47. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    ...If your poster material is nearing publication, consider uploading your work to a preprint server (and ... of the things we've mentioned above could be considered networking, but here are some more general tips...resume and prepare business cards. You might not consider being a student or postdoc a “business,” but having...

  48. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...batch analysis because of its ease-of-use and comprehensive reports. With CrispyCrunch, you get the all ...summarisation score".) For CrispyCrunch, we wanted to consider both factors, as a bench scientist would looking... our CRISPR 101 blog posts Learn about some considerations when designing gRNAs Find other CRISPR software...

  49. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...only help. Like any major life decision, the transition felt scary. While I knew two people from graduate...an industry postdoc is something that you’re considering. Like anything, there are pros and cons to postdocs... to enter academia, it is easier to make that transition from an academic setting—especially if job applications...

  50. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ... your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in cell lysis...equipment. Hopefully this post has provided some insight on these differences in viral production and will...versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses AAV and...
Showing: 401 - 450 of 881 results