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We narrowed to 849 results for: HA

Showing: 201 - 240 of 849 results

  1. Five Popular Model Organisms

    Type
    Blog Post
    ...more commonly known: the fruit fly. The fruit fly has been used in scientific research for over a century... a transgenic mouse line. Over the years the fly has become an ideal model organism to study an array ...development, genetics, and the nervous system. The fly has also emerged as an important human disease model ... described in 1993 by Norbert Perrimon’s lab and has been continuously improved upon since then. If you...Zebrafish (Danio rerio) Since the 1960s, the zebrafish has gradually become an important model organism. They...Caenorhabditis elegans) The worm, Caenorhabditis elegans, has been used extensively as a model organism since the...

  2. Evolution of Lab Techniques

    Type
    Blog Post
    ...abundance. If a scientist wants to know if an organism has a particular gene mutation, southern blotting can... DNA is complementary to the probe (for example, has the sequence of the mutated gene the researcher is..., two DNA fragments are observed because the DNA has the site where MstII cuts. In a patient with sickle...DNA fragment is observed. Today Southern blotting has been largely replaced by real-time PCR to answer ... large amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher...questions (see below). Importantly, although their role has changed, restriction enzymes are certainly not dead...

  3. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...refined to create Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing...example, Brainbow-1.0 expresses RFP when the construct has not undergone recombination. This default state was...neuronal pThy1-Brainbow constructs, Addgene also has two Brainbow adeno-associated viral vectors (AAV)...Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as ...neuromuscular junction. In zebrafish, the method has become very useful in lineage tracing; “Zebrabow”...Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section of...

  4. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ... first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns... low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently expressed...using chemiluminescent proteins for imaging, but it has precluded widespread adoption of the technology. ...Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing Nano-lanterns...

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

    Type
    Blog Post
    ...CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. It has the possibility to treat... applications.” Li’s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of... expanded organoids. “CRISPR in adult stem cells has huge potential, because the genomes of adult stem...

  6. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...for the flow plots discussed in this post, voltage has been adjusted appropriately and compensation performed...baseline expression of a protein while the green (top) has an upregulated expression (shift to the right). The...overlaps with the baseline suggesting this sample has a comparable expression to the baseline/negative ...allow for better interpretation. Priyamvada Prathima has a research background in protein mutagenesis and ...

  7. 5 Great Apps for Lab Life

    Type
    Blog Post
    ... safe to assume that anyone reading this article has a smartphone nearby. We tailor these devices to our...and a couple of other nice features. This app also has a great educational section. This section holds the...this MSDS app. The Chemical Safety Data Sheets app has a search function that allows the user to find an...memorized, but the structures are another story. AMINO has been very helpful when I’ve had to explain the properties...

  8. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...modified version of CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines. Cas9...effective in certain cell lines. This point mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro (PX459...that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction sites ...

  9. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...decades, recombinant Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the ...The most famous ones are influenza hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus...limitations of epitope tags and fluorescent proteins, it has been crucial to develop new tracers that enable accurate...are known to be notoriously difficult to label. It has also been shown that dendrites in the stratum oriens... provided better labels than fusions to 3 or more HA tags. 4. High-resolutions microscopy As indicated...

  10. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...technology has been studied and used in assays for over half a century; the Addgene blog has an excellent...new aptamer also has a higher melting temperature than its Spinach ancestors, and has a lower magnesium...initially still unstable in eukaryotes, the Jaffrey lab has since developed new RNA-based scaffolds to contain...

  11. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ... animal moving around to explore the environment has a different motivation (and brain areas associated... certain forms of drug addiction. When an animal has been exposed to morphine or other stimulating drugs...involved in locomotor sensitization? Previous research has shown that regions such as the nucleus accumbens ...dark compartment (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors... tests.    Forced swim test The forced swim test has been used since the 1970s to test the efficacy of...

  12. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ... in a town with a good public school system that has made strong efforts in the last 5 years to incorporate...curriculum. Our Parent Teacher Association (PTA) has been instrumental in supplementing that curriculum...some quick tips to get things going: If your PTA has a general meeting and a means to request funding ...without them! If you live in a school district that has a lot of parents in STEM careers, chances are they...nearly every talk I’ve given in the past 20 years has been aimed at grad students, postdocs, and other ...

  13. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...While much of CRISPR research has focused on genome editing, numerous discoveries have been made using...cleave the DNA. Much of the research using dCas9 has focused on transcriptional activation using a fusion...CRISPR-mediated purification of a specific genomic region has also recently been adapted to isolate genomic loci...tag systems including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification...for many other CRISPR techniques, this method also has great potential for improvements and adaptations ...

  14. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...altering the sequence of the protein. As DNA synthesis has become quite cost-effective it seems easy for scientists...optimize your gene of interest? Codon optimization has become extremely useful in expressing functional ...neurons in vivo (Slimko and Lester, 2003).  Addgene has a large selection of codon optimized genes, so check...which have even been linked to some diseases. It has been reported that 5-10% of human genes contain a...//doi.org/10.1093/nar/15.3.1281 Slimko EM, Lester HA (2003) Codon optimization of Caenorhabditis elegans...

  15. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors to light. One lesser-known...effector, and type IV with an unknown effector. Type I has subtypes A through G, with different compositions... Thanks to this complex, developing Cas3 tools has been challenging. It can be difficult to ensure comparable... job — Cas see DNA, Cas chop DNA. Cas3, however, has two jobs, one as a nuclease and one as a helicase...easier to work with than Cas3. Future targets Cas3 has the potential to be incredibly useful in therapeutics...

  16. The Challenges of Cell Culture

    Type
    Blog Post
    ...Addgene's Special Collections). Arguably, oncology has benefited most as cancer cells are amenable to culture...misidentified cell lines. The solution Fortunately it has never been easier to conduct high quality cell culture...identity. For example, perhaps a working cell bank has been made by a research institution (an excellent...at the scene of a crime (every human in the world has a unique STR profile) and can also be used as a paternity...topics. The LGC Group also provides standards and has laboratories to carry our independent quality control...

  17. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...firm The path to a career as a Grant Professional has been a long and winding path for many in the field...?  In the past 5-10 years, the Grants Profession has become just that, a profession. A grant professional...and the increasing regulatory process for grants has fostered the demand for experts in the field of grants... find your people Find someone who is working or has worked as a Grant Professional to mentor and coach...includes the social media community GrantChat. She has served in the nonprofit and government sector for...

  18. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ... in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes...protein interactions in living cells. Since then FRET has also been used to measure conformational changes,...brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP. The stoichiometry...Fluorescence lifetime imaging microscopy method FLIM has been developed more recently and is the most rigorous...emergence of faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...

  19. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    ...policy. Since early March, the open access publisher has required authors to include a note as to where readers...how much data was necessary and reasonable. PLOS has since clarified their data sharing policy and recently...Who is to say that the initial data-generating lab has efficiently trawled their own data? A distinct (perhaps...many have multiple, complex panels. PLOS, itself, has not finalized its definition of necessary data requirements.... Secondly, science is highly technical and this has a natural, albeit unintentional, tendency to work...

  20. Meet Aavery, Blugene's New Virus Friend!

    Type
    Blog Post
    ... and polling, Blugene's new virus friend finally has a name! We thank you for your participation in suggesting...play on viral vector terms. And, the name Kendall has a special place in our hearts: our first two offices...

  21. Scientist Networking: What is an Informational Interview?

    Type
    Blog Post
    ...Training as a scientist in the academic system has many pluses. I delighted in my graduate school years...call this building relationships because networking has gotten a bad reputation (as in…”I just hate networking...connections. A mistake made by many novice networkers has a potential to alienate connections. If meeting with...

  22. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...ElizabethKenkel) December 5, 2018 How social media has helped me in science It may have taken a bit of FOMO...Here are three examples of how using social media has helped me or my lab mates: For me, using social ...with social media as a scientist Using social media has enriched my professional development, and while it...helped you fit social media into your busy schedule. Has social media helped you advance your research or ...

  23. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    ...being targeted via RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly...sequence of the toehold switch. For example, Green has also developed toehold switches to detect Zika virus...protein motifs than previously thought. Sidhu’s group has made several hundred SH3 domain containing plasmids.... In addition to these constructs, the Sidhu lab has made hundreds of additional plasmids available through...

  24. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and variants ...scientific community. What are the features? CasPEDIA has collected relevant information on all the Cas enzymes...cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the other hand, generates...learn about Cas domains. CasPEDIA either directly has this information or links out to this content and...

  25. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...unsure of the next steps. Harvard PI Angela DePace has put personal development at the center of her lab... and postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine...

  26. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ... field of induced pluripotent stem cells (iPSCs) has been around for 10 years. In that time, scientists... of Myc or other oncogenes after differentiation has been linked to tumor formation in iPSC-derived and...iPSCs compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared to MMLV-derived...mechanism between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...cell division. A drawback to this approach is it has lower levels of efficiency at generating iPSCs, usually...reprogram cells is that it’s simple and efficient. It has a high efficiency of reprogramming when compared ...

  27. Cloning

    Type
    Guide
    ...DNA that already has restriction sites on either side of it into any plasmid that has the same sites in...clonase (creating an entry clone). The entry clone now has recombined attL sites flanking your DNA fragment ...fragment of interest. Now that your DNA fragment has been cloned into a donor plasmid, it can be rapidly shuttled...into a donor plasmid (or acquire one that already has your gene into it) and then using bacterial recombination..., such as restriction cloning, once your project has been completed via gateway cloning. Back to Top TOPO...pre-cut, linear, cloning ready TOPO® vectors that has a DNA topoisomerase I fused to the 3’ end. The topoisomerase...base pairs. Anything shorter than 200 base pairs has the potential to be completely chewed back by the...

  28. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts that can be assembled into...yeast MoClo is also for eukaryotes! The technology has been used to optimize protein secretion and expression...possible through MoClo. The MoClo Plant Parts Kit has everything you would expect in a MoClo kit plus the... general plant cloning experiments. Addgene also has two additional MoClo plant kits: Plant Parts Kit ...; Chamness et al., 2023). The Plant Parts II kit has a focus on infrastructure with shuttle vectors, several... gene silencing vectors. The Plant Parts III kit has a focus on transformation and genome engineering....Brychcy et al., 2023). For plant biologists, Addgene has not one, not two, but three CRISPR MoClo kits for...

  29. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... at the Georgetown University School of Medicine has used this gene editing system frequently over the...in human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor...via gene trap enrichment, whereas pAAV-TK-Acceptor has a conventional heterologous promoter-driven TK-neoR...editing compared to CRISPR editing CRISPR, of course, has started a revolution in gene editing. However, while... through aberrant double strand break repair, it has proven more challenging to develop high efficiency...

  30. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...reprogrammed less efficiently, no such correlation has been found between the age of the patients from whom...potentially promote tumorigenesis in vivo. The phenomenon has been recorded in past gene therapy trials and remains...reprogramming factors To date, the mRNA transfection method has not been used successfully to derive iPSCs from PBMCs...However, reprogramming of PBMCs by episomal vectors has yielded hiPSCs at a low efficiency (0.001-0.03%) ...efficiency: the pCXLE episomal vector toolkit It has been shown previously that suppression of TP53 enhances...

  31. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ... 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and ...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room for additional...the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type counterpart...amino acids, can also be packaged into AAV. NmeCas9 has the added advantage that it can be turned off by ...daunting, it’s actually a challenge that the field has overcome with split AAVs. In general split AAVs break...

  32. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ... maintain itself within host cells.  The plasmid has multiple ori that are functionally differentiated...various hosts (Pagatto and Dillon 2001).  The plasmid has its own replication initiation factors and proteins...the host. Host range depends on whether the host has the components to recognize the ori and allow for...restriction enzymes (Meyer et al. 1977). So if a plasmid has few restriction sites, it may be able to sneak past...commonly used bacteria in lab manipulations. Table 1 has a detailed list of the bacterial species these plasmids...

  33. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...quantified by cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-...performed on all new viral vector preparations. Addgene has used this method to generally validate our viral ... Material (AAV SFM), which is an AAV sample that has been quantified by 16 labs around the world and can...Hindson et al., 2011, Hindson et al., 2013), which has been shown to be more precise and more repeatable...2011): 8604-8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL,...

  34. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...challenge of keeping up on studies in many disciplines has been a long brewing frustration for me. I read a ...citations in their own work. Of course not all research has to be influenced by research in other disciplines...platforms were developing, Google acquired Youtube and has been growing its online video market ever since. ...this point most videos are based on research that has been published, but many discuss in-progress research...videographer, is a retired botanist and oceanographer who has a blog and several videos on sharing your research...

  35. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...University of California system, the postdoc union has negotiated a raise from $43,692 to $48,216, an increase...increase in salary of 58.3%. University of Delaware has a minimum of $36,360, requiring an increase of 30.6%...Hospital is the only part of the Harvard group that has responded to the FLSA at the time of writing, and...good thing. Reliance on a cheap trainee workforce has clearly not been a sustainable approach to biomedical...biomedical research in the U.S. and postdoctoral training has not been demonstrated to be required for the jobs...

  36. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...into Reddit. Here are some reasons why: 1. Reddit has a large and active community of people interested...right - the Reddit community is active, engaged, and has great questions! 2. The r/Science subreddit is well...the internet, using dialogue). Luckily r/Science has a great team of moderators who facilitated scheduling...preferred time to start answering questions. r/Science has pretty strict rules about not being too sales-y in...the requested topics to the Addgene blog The blog has many other excellent posts on science career topics...

  37. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use in mammalian...biology surrounding a particular gene (although it has been quite successfully adapted for use in screening... method for validating your selected gRNA’s, and has the bonus benefit of supplying you with primers for...sturdier than the delicate RNA and therefore there has less chance of contamination with RNase leading to...products will give specific details on what exactly has been inserted or deleted. Each lab will use a slightly...

  38. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...  Disadvantages Every rose has its thorns and every affinity reagent has its downsides. These reduced-in-size...created in lab in situations where a suitable antibody has been identified but a smaller or less immunogenic...using combinatorial libraries, and high specificity has been reported for a number of targets. Due to their...

  39. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...developing and optimizing biosensors in the lab. He has recently developed a third-generation intensiometric...release from hundreds of synapses at once. “This work has only been made possible due to the collaborative ... at the German Cancer Research Center (DKFZ) who has developed new redox sensitive GFP2 (roGFP2)-based...Qiu’s work on the PV neurons is in progress, she has validated the dual virus method to study CR neurons...

  40. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ... and serotype, respectively. For example, AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors... components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your gene...usually quite stable and in some mammalian tissues has shown expression for several years (Wonjo et al.,...scAAV vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) ...
Showing: 201 - 240 of 849 results