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Showing: 241 - 280 of 879 results

  1. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    Published
    Aug. 1, 2023, 1:15 p.m.
    ...don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same three nucleotides...Fidelity, it turns out, might depend more on the individual sequence(s) than on how well that sequence fits...Pryor et al., 2020), and randomly selected non-palindromic pairs allows for high-fidelity assembly for up...potential for mismatches, so you can redesign individual pieces as needed to improve fidelity. NEBridge...tool for you. Once you input a sequence, SplitSet finds optimal fusion sites to split it into multiple fragments...define the number of fragments, and the “search windows” you’d like the tool to use to identify fusion ... return the highest-fidelity overhang set it can find for those parameters. For even more specificity,...

  2. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...secondary structure to induce transcript cleavage, similar to the Rho-independent termination found in ...two general categories: rho-dependent and rho-independent. Rho factor is a helicase which assists RNA polymerase...common bacterial expression plasmids use Rho-independent terminators, which include naturally occurring... high-efficiency terminators such as T0. Rho-independent termination is also known as intrinsic termination...secondary structure and sequence conserved Rho-independent termination annotation for 90 bacterial elements...CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=39322032   No terminator is 100% efficient... than Polymerases I and II, which both require binding of termination factors. Although both are termination...

  3. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    Published
    Jan. 30, 2018, 2:42 p.m.
    ... all”. Manage the individuals in your team: learn what motivates them (keep in mind that it can be very...it will take time so be patient and keep an open mind. First let’s clear up a misconception about being...have reports with different levels of experience), find out what their learning styles are and capitalize...benefit of the team. By properly managing the individuals in your team you will have a much better chance...(needs to) feel valued. Talk to your manager and find a mentor Your manager should support you, just like... how to face a particular situation. But keep in mind that a good manager should help you work through...you and you need to encourage them to speak their minds. They will make mistakes and you will take responsibility...

  4. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...in the genomic locus. The first two breaks are induced via a generic PITCh-gRNA; the third break by an...flanked by 5' and 3' microhomology and PITCh-gRNA binding sites. This PITCh plasmic is cotransfected with...optimized the PITCh-gRNA for minimal off-target binding in mammalian genomes as assessed by CRISPR design... and 3’ junctions, respectively. These results indicate that PITCh is a robust method for genomic insertion...staggered pattern, it is thought to be ideal for HDR-independent knock-ins, but this possibility is still being...systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from this publication at Addgene. ... Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids from this publication at Addgene. 3. Zhang...

  5. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ... of binding DNA to silica columns, it’s bound to glass syringe filters. Normally DNA does not bind silica...silica are commonly used to bind and purify DNA, cellulose also efficiently binds, or at least entraps nucleic... can be stacked together in series to increase binding capacity. Figure 1: Purify DNA using glass... and 2) it helps DNA stick to the glass. After binding DNA, the glass particles are pelleted by centrifugation...mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids, so placing... Addgene blog Read our Plasmids 101 blog series Find other plasmid protocols and tips Browse our plasmid...plasmid cloning blog posts Resources on Addgene.org Find protocols for molecular biology and cloning Read...

  6. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...the tTA inducer virus, but the desired sparsity of labeling is achieved with lose doses of inducer virus...Gradinaru Lab at Caltech stepped up to the challenge. Find the PHP Plasmids Here! Their Solution: Use the ...viral cocktail are injected, by chance lots of individual cells will be transduced with multiple viral ...notice in figure 4b how difficult it is to trace individual cell’s axons, particularly the two green neurons...expressing RGB proteins under the control of an inducer, i.e. tet-off transactivator (tTA), is injected...that are labeled. Part 2: An AAV expressing the inducer (i.e. tet-off transactivator) is co-injected. The...tag-then-express system that allows for controlled induction of multi-color labeling of neurons (figure 4)....

  7. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...Addgene.org Find CRISPR plasmids and resources here Find pooled libraries available at Addgene Find CRISPRko...mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to a repressive... and Doench lab selected sgRNAs in this optimal window and ranked them based on their optimized sgRNA ...Brunello knock-out library while mitigating toxicity induced by high copy number dsDNA cutting. A Dolcetto library...transcriptional machinery to dCas9 and its RNA directed binding site. These methods include the fusion of an activation...overlapping genes, most genes were unique to the individual libraries. These unique hits may have been false...can be used in complement to study gene function. Find the Calabrese pooled library and lentiviral prep...

  8. Editor's Choice, September 2016

    Type
    Blog Post
    Published
    Oct. 7, 2016, 4 p.m.
    ...describes a technique you can use to find phage proteins that bind to your bacterium of interest. For those...contributed the most to this record breaking month and to find other posts that deserve a second look. 5 Great ...colleagues. Technique: Probe Phage Genomes for Host Binding Proteins & Running for Rare Disease, Running for...first, "Technique: Probe Phage Genomes for Host Binding Proteins," was contributed by guest blogger Jessica...dress up as Mr. Incredible, an added bonus. You can find the rest of our September posts in chronological...CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats Better Dyeing Through Chemistry... a Lab and a Project That’s it for now. As I indicated in the opening paragraph, I’m currently on my ...

  9. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...Blog Read other Addgene news Find the most recent plasmid blog posts Find the most recent CRISPR blog ...Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary Valley,... Jennifer Tsang There are many strategies for finding your clones in an experiment. Read this blog post... Learn about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at...are nucleic acid sequences that fluoresce upon binding a small molecule, are often named after vegetables...Tools for Making Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for making scientific...posts Resources at Addgene Browse the repository Find educational resources at Addgene Search for plasmids...

  10. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...post-meeting survey indicated that the program was largely a success, but also helped us find areas for improvement...really fit in anywhere and everyone who was doing the kind of work my lab wanted to do already seemed to know...Singletons came together. The term Singleton was on my mind because we’d had discussions in my lab about whether...Image: James et al., 2020. Help me stranger - Finding mentors for the meeting Just like analysis pipelines...genes that can improve chances of survival for an individual, and eventually, a community. The experiences...and vastly expand their networks as they naturally find people with which they have interests and experiences...experiences in common. We often refer to this as “finding your people.”    Had a fun time at the #BinningSingletons...

  11. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    Published
    Jan. 23, 2020, 2:45 p.m.
    ...have the same blind spots, and there wouldn’t be anyone to question these blind spots. Blind spots often...development is not one of the things that comes to mind. The truth is, there is a lot of software development...result of our diversity which in turn minimizes the blind spots in our work.  Agile at Addgene We follow an...short periods of time called sprints. The team is mindful of the number of stories we commit to for each ...reviews because it’s like a little peek into the mind of the developer and I get to see an approach that... this happens, we need to research and hopefully find a comparable library if it’s gone stale or we will...Learn about what went into our search engine update Find all Addgene news Resources on Addgene.org Check...

  12. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...on Addgene.org Find the Spaghetti Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV...brain. This atlas enables researchers to quickly find the best AAV serotypes to target the neurons they...synthetic peptide FLAG, the synthetic streptavidin-binding strep-tag and, more recently, OLLAS (Escherichia...composed of FPs fused to multiple epitope tags. Indeed, to create the smFPs Looger’s team strategically...a physics graduate from Oregon State University. Find the Spaghetti Monster probes at Addgene Uses of ...structures through their multiple, high-affinity binding sites for primary antibodies which can be used ...Through their multiple, highly specific antibody binding sites and primary antibody-based detection, smFPs...

  13. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...hoping to follow the traditional academic route (find tips for getting a faculty position here). Possible...comprehensive lab experience. With this valuable training behind you (and largely only that is necessary), there...problems, a general positive attitude, and an open-minded personality. These traits will help you deal with... a customer’s concerns and are patient enough to find the real issue, you will do well in a technical ...troubleshooting or issuing replacement products. In order to find good and helpful answers for customers, internal...are often found in sales or lab-based roles in industry. What to look forward to in a technical support...fascinating and exciting to listen to the many individual stories and get (sometimes refreshing) insights...

  14. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ...bARGSer with several inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and ...replication (p15A) worked best under a wide range of inducer concentrations (0.1-1% L-arabinose) without signs... or at lower temperatures for longer. Cultures induced with arabinose and colonies on arabinose-containing...expression, from left to right: colony whiteness on inducer plates, white inclusions in cells with GVs via ...integratable cassettes, both with doxycycline inducible promoters (Fig. 2a). The first cassette contains... phase contrast microscopy just one day after induction of expression… but longer expression leads to ...plasmids (Fig. 2b). Next, perform a short (12-hour) induction with doxycycline prior to FACS, gating for the...

  15. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...within the editing window (bases 4-8 of the protospacer). While this is a small window, this system can ... website Find CRISPR Plasmids Catch up on your CRISPR background with our CRISPR Guide Find Software ...through Addgene’s repository) and make it easy to find your perfect match. Choosing a Cas9 for your knockout... anywhere to be found. If you’re lucky enough to find a SpCas9 compatible PAM at your site of interest...performed on all relevant bases within the editing window, for example all C’s will edit to T’s. Thus, it... capable of interacting with an additional RNA-binding transcriptional activator. Plasmids for activating...the many freely available sgRNA design programs. Find Validated gRNAs for Your Next CRISPR Experiment ...

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

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the power of ...thanks to our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD ... long distance runner, gardner, and cook.      Lindsey M. Costantini received her PhD in Biomedical Research...Pubmed Central PMCID: PMC557822. 4. Costantini, Lindsey M., and Erik Lee Snapp. "Fluorescent proteins in...Pubmed Central PMCID: PMC3806368. 5. Costantini, Lindsey M., et al. "A palette of fluorescent proteins optimized...Central PMCID: PMC1222268. 7. Aronson, Deborah E., Lindsey M. Costantini, and Erik L. Snapp. "Superfolder ...Pubmed Central PMCID: PMC3359384. 9. Costantini, Lindsey M., et al. "Cysteineless non-glycosylated monomeric...

  17. 10 Ways to Share Your Science!

    Type
    Blog Post
    Published
    May 13, 2014, 8:34 p.m.
    ...help when it comes to finding a job. Many people in science will work in industry or take on collaborative... way to follow news about particular subjects or find out what others are reading. Check out science blogs...tell your friends or others in the field where to find it. A good way to reach others in the science blogosphere... 5)   Local science museums are a great place to find opportunities for science outreach. Here in the ...student I know who participated was surprised to find that his friends and family finally understood why...office is a good place to start. It’s not unusual to find science writers who started out as graduate students...they’d be willing to meet for lunch or coffee and find out how they got where they are. 8)   Look into ...

  18. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators NIR-GECOs...researchers choose the probe for their needs. You’ll find basic information on NIR FPs, their properties and...of state-of-the-art probes, many of which you can find at Addgene. Biliverdin chromophore Near-infrared...listed in Table 1) were optimized for efficient binding of endogenous BV.  When imaging NIR FPs, consider...performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova et al., 2021...encoded near-infrared fluorescent calcium ion indicator. Nat Methods 16, 171-174 (2019). Matlashov, M..... A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol 39, 368-377...

  19. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ...Repair (HDR). NHEJ is the canonical homology-independent pathway as it involves the direct ligation of...synthesis-dependent strand-annealing (SDSA), and break-induced repair (BIR). The non-conservative method is single... synthesizes DNA from the second DSB end. The individual HJs are resolved via cleavage in one of two ways...to fill the gap, thus completing SDSA.   Break-induced repair (BIR) pathway BIR is characterized by the...pathway may not be immediately applicable in DSB-induced genome engineering, it has biological importance...as 30-50 bases for each homology arm, but keep in mind these numbers may vary based on your locus of interest...double-strand break repair and their potential to induce chromosomal aberrations. Pfeiffer P, et al. Mutagenesis...

  20. Summer SciComm Series: Master of SciComm

    Type
    Blog Post
    Published
    July 21, 2022, 1:15 p.m.
    ...entails. Despite being engrossed by the theory and findings of brain science, my curiosity did not ultimately...communication. The first year of the course involved a whirlwind of skill acquisition. Classes covered filmmaking... would have a high level of career satisfaction indeed.   For many venturing forth from tertiary study...discernible paths. The first leads into the world of industry. Along this road, science communication graduates...Moreover, this subject allowed me to engage in findings relevant to my background (neuroimaging, biomechanics... a survey instrument designed to measure how individuals respond to issues of unsettled science. Understanding...building, each day brushing past and chatting with individuals whose research ranged from wildlife conservation...

  21. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    Published
    June 17, 2021, 1:17 p.m.
    ... can find all the plasmids deposited from Di Ventura's lab, including the popular light-inducible nuclear... is about a new tool called BLADE (blue light-inducible AraC dimers in E. coli) that turns expression ...promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC to create BLADE...exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain, Vaucheria frigida aureochrome1...tested different linkers between VVD and the DNA-binding domain of AraC using a mCherry reporter downstream...Adam." This bacteriograph was created using 160 individual images stitched together. Image from Romano et...resources on the Addgene blog Learn more about inducible promoters Visit the Plasmids topic overview page...

  22. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    Published
    Aug. 25, 2015, 2:30 p.m.
    ...  Your mind is likely drawn to the Mark Zuckerbergs and Evan Spiegels of the world – individuals in their...contribution to the healthcare landscape.  You will find an extra layer of difficulty launching a company... You've spent your life making connections and finding mentors -- well, it's time you call upon these ...hurt, but if you pitch to enough people, you will find the answer to your viability question. The miLab...Build a team of 2 or 3 Don’t go it alone.  You’ll find that no matter how well you know your idea, you ...science space if you don’t have an advanced degree behind your name, but don’t let this deter your [email protected]       Additional Resources Find more career posts on the Addgene blog Read our recent...

  23. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ...expression, turning transcription on and off by binding proximal to their target genes. Since gene expression...), barcoded for tractability. These ORFs are individually overexpressed and available for lentiviral packaging...large sequencing endeavor, a map of possible TF-induced cellular states was generated (Joung et al., 2023...association of the uncharacterized TFs is a useful finding. Transcriptional program changes from this screen...identify both up- and down-regulated pathways. Individual cellular fates were then matched with candidate...pooled library. Louvain community clusters of individual cells along with color coded nominated reference...fluorescent controls. Alternatively, you can order individual plasmids from the library to further investigate...

  24. Management for Scientists: Delegating is Key

    Type
    Blog Post
    Published
    Feb. 20, 2015, 8:30 p.m.
    ... together teams of individuals who enjoy and are experts in each of the individual tasks required to develop... but through the coordinated efforts of many individuals. Check out Joanne's Reddit AMA   Benefits of...each component of the project completed by an individual who is a local expert in that task (or has the...outcomes clear? If you don’t have clear outcomes in mind, you won’t be able to communicate these well to ...articulate the desired outcomes. Begin with the end in mind. Specify desired results. Are you delegating to...Set Concrete Goals with Your Mentor through an Individual Development Plan Resources on the Addgene Website...Website Are You a Molecular Biologist? Find Plasmids for Your Research Here!        ...

  25. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    Published
    Nov. 6, 2015, 3:30 p.m.
    ...instance there are few resources to help get individuals started on the path to internationalization and...development and training in a globalized world where individuals can exchange ideas, network, collaborate, and... supportive and protective environment where individuals are empowered to productively navigate through...others have faced yet, still, this challenge did not hinder them from advancing their careers and achieving...skills, positive disposition and motivation, open mindedness, and the ability to accept feedback and adapt...to 78 undergraduate/graduate students from 5 independent universities in Spain through 22 international...Country. Mentees represented a diverse group of individuals with technical and career path needs, and came...

  26. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...optimized with zebrafish in mind, it can easily be applied to other systems.    Find BLITZ Plasmids here! Xiong...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...deposit please sign up here. Here's what you'll find in this post: Improving flexibility for in vivo...GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target stabilizes the...mouse. Image from Goertsen D, et al., 2022.  Find M-CREATE plasmids here!  Goertsen D, et al. Nature...microbial SSAPs. Image from Wang, et al, 2022.  Find dCas9-SSAP plasmids here! Wang et al. Nat Cell Biol... (TRE) or kinase translocation reporters (KTRs). Find visual barcode plasmids here! Kaufman, et al. 2022...

  27. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    Published
    June 29, 2021, 1:15 p.m.
    ..., you may be curious about all of the different kinds of antibodies that are available. One common type..., a limiting dilution is performed to isolate individual hybridomas and ELISA is used to identify the ...useful when you want to ensure that the antibody binds to a specific region of your protein of interest... example, if you are studying a protein that may bind a ligand during your experiment, then you’d want...that recognizes an epitope far from the ligand binding site (Mould et al., 2016). If your antibody recognizes... monoclonal antibodies will exhibit off-target binding to unintended proteins. This is because monoclonal...between lots over time. Specifically, the antigen-binding sites of the antibody - the paratopes - may change...

  28. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...of proteins (iLID and SspB) that undergo light-induced dimerization. Upon application of blue light, iLID...a conformational change allowing high affinity binding to SspB, brings together the split botulinum neurotoxin...that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al...segment In recent years, scientists are beginning to find connections between the microbiome and diseases,...systems that are potentially problematic for imaging. Find the ARG plasmids at Addgene. For more details on...multi-input cell classifiers in one assembly step. Find the miRNA reporter collection at Addgene! Gam JJ...

  29. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...nicknamed CasRx), as well as EsCas13d. Find the Yan et al. plasmids here Find the Konermann et al. Plasmids here...Cas13d orthologs described in these papers can independently process their own CRISPR arrays into guide RNAs...cleavage is retained in dCas13d and is thus HEPN-independent. These enzymes also do not require a protospacer...RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader...even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable...Since catalytically dead dCasRx maintains its RNA-binding properties, Konermann et al. tested its ability...C2c2) Characterization Resources on Addgene.org: Find RNA Knockdown Plasmids Check out our CRISPR Guide...

  30. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...implemented and how they’ll make it easier for you to find the plasmids you need. We'll be monitoring and improving...and effective changes will give you the ability to find the plasmid with the features you need quickly. ...details. Click on the “View all sequences” button to find all available sequence information from Addgene ...hover over the ampicillin resistance gene, you’ll find the full name of the gene, the name of its gene ...functionality. Restriction Enzymes Here you'll find a table of restriction enzymes that cut the given... times they cut. Use the filters on the right to find single, dual, and 3+ cutters. You can also sort ...feature names, locations, sizes, colors used to indicate them on the map, direction (if relevant), and ...

  31. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ... All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein...Fluorescent imaging techniques have become indispensable tools for molecular and cell biologists over...sensor displaying a 500% signal change upon calcium binding. Suzuki et al. also envision eNLs as good reporter...used a linear unmixing algorithm to decode the individual colors. The development of this algorithm is ... (2012): 1262. PMID: 23232392 PMCID: PMC3535334 Find plasmids from this paper at Addgene. Takai, Akira...2015): 4352-6. PMID: 25831507. PMCID: PMC4394297 Find plasmids from this paper at Addgene. Suzuki, Kazushi...(2016): 13718. PMID: 27966527 PMCID: PMC5171807 Find plasmids from this paper at Addgene. Additional...

  32. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    Published
    June 4, 2015, 1:03 p.m.
    .... If you are interested in reading more, you can find the rest of the Plasmids 101 posts here. Now that...β-galactosidase. Expression of the lac operon is induced by lactose, and also by a lactose analogue, IPTG...thiogalactopyranoside). (To be completely accurate, IPTG binds and inactivates the lac operon repressor, thereby...dye-linked substrate called x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble... insoluble blue pigment (4-chloro-3-brom-indigo). So far, so good, but how does this help us screen plasmids...insert. All colonies on this plate should be blue, indicating that your IPTG and x-gal are working as they ...of false positives: Blue-white screening only indicates the presence of AN insert, not necessarily YOUR...

  33. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    Published
    July 10, 2014, 7:17 p.m.
    ... initially include a visionary, a science-minded individual, and a specialist. Notably, the team must ... Bruce Booth How to Find a Job in Biotech and Resume/CV tips - LabSpaces Blog Find a Buisness Plan Competition...into a startup company. However, like many like-minded people, I quickly realize that a company requires...transitioned away from the bench and became all about finding the best product-market fit for this set of technologies...is. While communication styles in academia and industry vary drastically, he reassures that one can learn...anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes. The ...

  34. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...channels, the team will be able to validate their findings in mammalian cells with both human and murine ...cytokines actually get out of keratinocytes and induce their effector functions. It’s an important process... Hospital Zurich. IL-36 cytokines are a potent inducer of an autoinflammatory loop: keratinocytes both...Lin got me interested in TS when I was a newly independent faculty member studying sex differences in thoracic...thoracic aortic disease,” Prakash says. In induced pluripotent stem cells derived endothelial cells from...thirty GNB1 missense variants identified from 58 individuals with GNB1 disorder since the identification of...allow us to enhance our biosensor repository as we find ourselves lacking several crucial biosensors that...

  35. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    Published
    Feb. 2, 2018, 3:20 p.m.
    ...You need a lot of time to let you mind process things. Sometimes finding that uninterrupted time to be creative..., but started looking for a new job because “it kinda sucked to work remotely. I was also tired of the...integrated and part of a team.” It took her two years to find her current position at Fred Hutch because her skill...through the tough assignments, the skills that Susan  finds important for her work include: “Listening. Evaluating...advice for getting started in science communication: “Find as many opportunities as you can to get your name...unpaid work as a student. For post-grad school individuals, Susan recommends starting a blog, but with one...internship. Application information can be found here Find tips for using social media as a scientist here ...

  36. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ...with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...endogenous ERK, and its localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates...Researchers commonly use rectal bacterial infection to induce tail swelling in C. elegans, but activation of ...cells by 24-48 hours exposure to blue light also induced tail swelling in ~75% of worms (see below figure...photoswitchable kinases in my experiments? You can find the plasmids from Zhou et al here. In addition, ...empty backbones in the Michael Davidson Collection Find Photoswitchable Fluorescent Proteins ...

  37. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...to autophagy-inducing conditions. Rosella makes it easier to study the mechanisms behind autophagy by... cells. Autophagy (Greek for “self-eating”) is induced by a lack of nutrients and targets cytosol and ... via proteasome degradation. Autophagy is then induced and quantified by measuring the amount of radioactivity...radioactivity in the culture supernatant (this is indicative of protein degradation). To control for protein...cytosol in an inactive form unless autophagy is induced. Then non-selective macroautophagy of the cytosol...some conditions it aggregates in an autophagy-independent manner. Additionally, tracking Atg8p or LC3 doesn...fluorescence. After 4 hours of nitrogen starvation to induce autophagy, red but not green fluorescence accumulates...

  38. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...version, but if you find yourself doing your own conjugation, it is important to keep in mind that you will ... be useful. Mainly because, well, antibodies are kind of hard to see. To solve this issue, researchers...antibody conjugates in an earlier post, where you can find some additional tips for working with these tools...affinity for avidin and streptavidin, both of which can bind up to four biotin molecules and can themselves be...with significant multiplexing potential. You will find antibody-oligonucleotide conjugates used in assays...protein-protein interactions with other peptides, like an Fc-binding domain (Figure 1).  Figure 1: Examples of...). If some of those lysines are in the antigen binding region or even just nearby, then conjugation could...

  39. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ..., coughing, etc. Also, a constant in-flow of individuals wherever the cell cultures are kept will increase...Contamination! Tissue culture media often have indicators included in the media that turns yellow from ...trigger a series of chemical reactions and this induces a noticeable color change. This method has high...fluorescent signals produced by these bacteria can hinder correct result interpretation. Also, the signals...Also, prolonged use of a particular antibiotic can induce resistance for other bacteria apart from mycoplasma... should be cleaned regularly and dampness of any kind should be avoided. Distilled water provided to maintain...because you are not the first person to face this kind of problem. Educate others when you can and help...

  40. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...that absorb photons for energy conversion or the induction of intra- or intercellular signaling pathways ...extra-chromosomally long-term expression and low ability to induce innate immune responses (Fenno et al., 2011). However...site-specific recombinases such as Cre or Flp that induce DNA recombination at defined recognition sites ...between mismatched recognition sites, its presence induces first opsin inversion, and then lox sites excision...opsin gene; in Cre expressing cells, however, Cre induces the recombination of the double-floxed opsin construct...FLEx-based vectors to study the neural circuitry behind the mechanisms of sleep and anesthesia. In Cre ...that it is possible to separate neuronal inputs to induce aversion- or reward-related behaviors. The high...
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