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We narrowed to 871 results for: sta

Showing: 181 - 200 of 871 results

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

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...single cell isolation and downstream sequencing (Standage-Beier et al. 2019). TREE is a real-time, fluorescent...in emission spectra from BFP to GFP. Image from Standage-Beier et al., 2019. The team put TREE to the...reporter proteins. Protein translation requires the start codon of AUG immediately downstream of a kozak sequence...mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image adapted from Katti et al., 2020...generated a silent GFP construct, that contains an ACG start codon mutation, and integrated it into human and...As the initiation of protein translation at the start codon ATG is universal, the Dow lab successfully... including mScarlet-I, luciferase, antibiotic resistance markers, and other enzymes such as Cre-recombinase...

  2. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ... laboratory tool often employed to establish cell lines that stably express a gene of interest. While ...background of the lines you are using, it is best to start from scratch. Obtain a vial of the desired cell ...production and infection.     Great transduction start with great virus If having happy cells is the first...extreme changes in osmotic pressure and pH can destabilize the viral envelope. One drawback of the multiple...The effect of freeze-thaw cycles on lentiviral stability depends on a variety of factors such as the transgene...al. found that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, ...prep!) and infect. While this approach is a great start to a successful infection, one point to consider...

  3. An Addgene Co-op: The Intersection of Fun, Friendship, and Work

    Type
    Blog Post
    Published
    June 29, 2023, 7 p.m.
    ...special about her pure interest and curiosity to understand my work and me, especially as much of her focus...environment filled with people who are interested in understanding the work you do, in addition to being genuinely...bigger projects I was tasked with was creating and sustaining a biweekly report of our social media analytics...Addgene before. This meant I was responsible for establishing a new report and routine across the marketing...excited but overwhelmed; I had no idea where to start. Thankfully, my managers set aside time for me to...template to build out a report on. I was able to understand what metrics would be useful and how to organize...with the larger content and marketing team, and I started preparing for my first official professional presentation...

  4. Management for Scientists: Managing vs. Leading

    Type
    Blog Post
    Published
    March 31, 2015, 4:08 p.m.
    ...work gets done Managers do this by removing obstacles that stand in the way of the doers and by ensuring ...just have to know it when you see it? “The Three Stages of Your Career: Doer, Manager, Leader” by Charlotte...importance of these very different roles. At any stage in our careers we may be in a cycle of doing, managing...are the ones who do the work Doers focus on understanding what is expected of them and learning new skills...look to the future Leaders must get new things started and then step back to let others take the reins...productive culture and to correct systemic problems. Understanding these three roles allows you to identify what...the requirements of the current task. Most of us start as doers, some become managers and some take on ...

  5. The Strength of Story Telling

    Type
    Blog Post
    Published
    Dec. 19, 2017, 2 p.m.
    ...conclusion. Advice for Telling a Good Story Start at the start Any good story needs a solid point to kick...at. Whatever you’re working on, start wherever it makes sense to start and try to be as chronological ...almost all of us are programmed to appreciate and understand. We build narratives around the self, too — ...author. When I raised this, he was pretty angry and started arguing with me. First I calmed the situation down...for all of us.” This is a classic example of the STAR (Situation, Task, Action, Result) interview technique...and engage with and what parts you can leave out. Start viewing your work in the form of a story, and you...telling it. End at the end A story needs a good start point and it needs a good end point. One thematically...

  6. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ...conformational states: bent-closed, extended-closed, and extended-open. In the two closed states, integrins...its open state, activating the receptor. Ligands can bind to integrins in their closed states (A) and ... ligands with high affinity. This extended-open state is key to strengthening ligand binding, mediating...in the extracellular environment. This further stabilizes the extended open integrin conformation. During...B) but with much lower affinity than their open states (C). Figure courtesy of IPI. IPI anti-integrin...rat or mouse variable regions fused with rabbit constant domains. All bind specific integrin alpha subunits...recommended for use in flow cytometry. Based on the established applications of the original hybridoma monoclonals...

  7. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    Published
    Jan. 4, 2022, 2:15 p.m.
    ...feelings, low morale, poor decision making, and stalled projects. You want to identify and address these...also be categorized as either “personal” or “substantive”. A personality-based conflict occurs when two...a lab or work environment if left unchecked. Substantive conflict occurs when people or teams who normally...”) when in fact many sources of conflict are substantive (“Person C told Person A one thing and inadvertently...in my current role revolve around missed tasks, stalled projects, and disagreements over prioritization...through multiple channels is not reasonable or sustainable. That said, when you know that a person or team...can indicate confusion around... Ownership: Misunderstandings about who is responsible for what tasks or...

  8. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology...interference is known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding...Organized Smooth Endoplasmic Reticulum (OSER) assay (Costantini et al., 2012) measures homodimerization of FPs...This assay is quantitative and is currently the standard for assessing oligomerization in cells. In addition...protein characteristics such as the brightness, photostability, maturation time, and acid sensitivity of various...parts of the fusion from N- to C-terminus. For instance, Lifeact-mTurquoise2 (Plasmid #36201) or mScarlet-tubulin...blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...

  9. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...post-translational modifications, identifies a staggering ~250,000 phosphorylation events in the proteomes...proteomes of higher mammals (2). How can we begin to understand the importance of any of these phosphorylation...not reversible, and thus cannot reflect the true state of protein modification within the context of its...difficulties with kinase specificity in vitro. The establishment of a system in which purified, recombinant protein...provides researchers with a valuable tool for understanding the contribution of phosphorylation to enzyme...and second, through a gel shift assay using the PhosTag system (see below). This plasmid backbone can also..., one should optimize protein production using standard protein purification strategies; expression should...

  10. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    Published
    Nov. 25, 2015, 4 p.m.
    ...postdoc salaries as opposed to Staff Scientists Increase the Starting Salary for Postdocs to $50,000 ...symposium will prove incredibly important to the stability of the biological research enterprise going forward...need to know in order to even assess the current state of research became more clear throughout the various...information about the postdoc work force, problems start with the question of “What is a Post-doc?” The National...cost-effective for PI’s to pay postdocs as opposed to staff scientists. This results in a glut of postdoctoral... De-incentivize going into PhD programs as the standard option for scientists by requiring graduate students...$50,000 (the current median postdoc starting salary as reported by Paula Stephan is $40,000) Increase Knowledge...

  11. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...Macaques N/A Stanton et al., 2023 AAV-MDV1A CNS Mice No (LY6C1) Stanton et al., 2023 ...The blood-brain-barrier (BBB) exists to prevent substances in the blood, like proteins and viruses, from...into the brain. While helpful from a biological standpoint, it makes delivering AAVs to the brain difficult...interest, and health. While it’s always helpful to start with a literature search to see what others have...considerations you’ll need to be aware of before starting an experiment. Controlling expression As systemic...sensors, CRISPR elements, etc... *These are only starting recommendations; you will still need to experimentally...are a good fit for your experiment, it’s time to start selecting and testing systemic capsids! Figure 2...

  12. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...Kostman.   Important considerations before you start an experiment with CRISPR The hammer, the jigsaw... an alternative ATG downstream of the annotated start codon. Likewise, we avoid target sites that code...undertaken lightly! Indeed, truly achieving the gold standard requires not one but two rounds of single cell...in a 5 - 10 nt window relative to the PAM, and bystander edits are possible if there is another target ...target a ~100nt window upstream of the transcription start site (TSS), while for CRISPRi, a ~100nt window downstream...the target (e.g. accessibility due to chromatin status). No computational prediction is ever perfect, ...nevertheless still lead to activity has been established, and can be used to identify sites that are likely...

  13. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...recognize the constant region. Nonspecific binding, caused cell receptors for the constant region, is also...about an affinity reagent, they’re talking about a standard antibody: An ~150kD protein with both light and... consistency in the final product. Because the constant (non-epitope binding) region is the first thing...connected by a short linker peptide. The lack of constant region makes them less immunogenic, but they have... protein nanostructures. Diabodies readily co-crystalize, a major advantage over antibodies (Kwon, 2019...signals due to their monovalent nature. They are stable up to 80°C and can refold correctly after denaturing...domain as their scaffolding instead of the antibody constant region. They are generated using combinatorial...

  14. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ...antibodies). By the end, you’ll have a better understanding of what makes polyclonal antibodies unique and...protein. Whereas production of monoclonal antibodies starts with selecting just one of these B cell clones ...immune response. Once the initial antibody response starts to wane about a month later, the animal is given...target protein. While this can increase background staining (see next section), it may be advantageous if ...protein in low abundance or whose conformational state may change. If your protein might become slightly...greatly increasing signal amplification.  When to stay away from polyclonal antibodies The most prevalent...might experience variability in the strength of staining and background noise. If you do use different ...

  15. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    Published
    Jan. 11, 2017, 3:30 p.m.
    ...$47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was due to the threshold at which...to now - across a wide consensus of academic stakeholders. However, while most institutions are indeed...the lapsed mandate (or are being compelled at the state level, in the case of some public institutions) ...do so. Immediately after the injunction, as we started to hear of institutions cancelling plans to raise...their raise were: postdocs at institutions like Stanford and the Whitehead Institute, where salaries had...continuing to reach out to institutions with a statement indicating that we encourage them to keep raising...across geographical regions When we originally started gathering this information, many assumed that those...

  16. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    Published
    Oct. 1, 2019, 1:21 p.m.
    ...academic scientists are starting to see the advantages of using social networking to start, build, and track...tell when some start-ups were about to be bought by seeing certain groups of people start to update their...my seven best tips beyond the basics to get you started. 1) Be visible and open to connections This is...add a personal note, you waste an opportunity to start things out right. Also, many people on LinkedIn ...include news, inclusion, culture and connections. Start your interview research there! 6) Your LinkedIn ...good or better than your resume Don’t make the mistake of having a shoddy profile and not updating it....changes. If you are working, and all of a sudden you start updating your profile, everyone will think you are...

  17. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...increases when target DNA strand separation is stabilized (2,3). Stable strand separation is maintained by 2 sets...HF1 each stall in an inactive form after binding to a target site and propose that this stalling leads to...biological disciplines, the base technology itself is constantly improving. Cas9 variants have been modified for...and the negatively charged DNA and therefore destabilize basal strand separation. This decreased strand...separation and target DNA binding to the Cas9 nuclease. Stable strand separation is maintained through interactions...mutations in the mobile Rec3 domain facilitate this stall. With this new found structural knowledge, in collaboration...publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal structure of Cas9 in complex with guide RNA and...

  18. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...MMLV-derived Retrovirus Integrating High Low Efficient and Stable Genome integration Insertional mutagenesis, transgene... Lentivirus Integrating High Low Efficient and Stable Genome integration Insertional mutagenesis, residual... vectors: This approach is the same as using a standard lentiviral vector, but transient expression of...for the oriP/EBNA1 plasmid to be maintained as a stable extrachromosomal piece of DNA. Reprogramming efficiency...large nucleoli, and little cytoplasm. Positive staining for alkaline phosphatase, expression of embryonic... the ability to generate teratomas is the gold standard instead. Teratoma formation requires injecting...two categories: 1) studies focused on better understanding the mechanisms of reprogramming, and 2) studies...

  19. Plasmids 101: Modular Cloning

    Type
    Blog Post
    Published
    March 12, 2024, 1:15 p.m.
    ...Hurt. Understanding how individual parts can be assembled into TUs is key to understanding MoClo. By...another optimized platform. Gene Stacking As the name implies, gene stacking is the use of a combination of...transcriptional units, and multigene constructs. Let’s start by examining individual parts and transcriptional...plant biology to achieve traits such as disease resistance. MoClo is a great tool to deliver many combinations...MoClo Interested in using MoClo? You don’t have to start from scratch! As you can imagine, there is an array... many different MoClo kits have already been established. Keep an eye out for our next blog on choosing... Marillonnet S. A modular cloning system for standardized assembly of multigene constructs. PLoS One. ...

  20. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ...connections to the starter cell, RABV was modified to only infect user-defined starter cell types, as well...’s architecture and plumbing are important. We started with learning the head is connected to the neck...the whole brain, it first infects one cell (the “starter cell”). It then replicates within that cell, and...retrogradely infect neurons that project to the starter cell. Once this first order infection occurs, subsequent...). Based on this, cell type specificity of the starter cell can be achieved by using EnvA-pseudotyped ...selectively supplying the G protein in trans only to starter cells (infected with G-deleted RABV). In this case... case, G-deleted RABV will infect the starter cell. RABV replication will then occur and the G protein...
Showing: 181 - 200 of 871 results