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  1. Advice for Choosing a Research Project

    Type
    Blog Post
    ...interests. We've outlined some important factors to consider and summarize helpful advice from the successful...Achieve Success in Academia". Important factors to consider when choosing a research project After joining...and goals. The following questions can serve as a guide when selecting a research project. What will the... are available in order to realize your project ideas. What mentors are available for guidance, intelligent...fungal biology.” Connie Cepko advises students to consider their goal of PhD training when selecting a research... good judgement with the current technology to identify a problem, asking what approach will I take and...touching a pipetteman. Take the time to generate new ideas and think about them critically before diving in...

  2. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ... nucleases from re-identifying it (3). Surprisingly, few genes have been identified that neutralize CRISPR...Addgene’s CRISPR Guide or find CRISPR/Cas plasmids for your research. Do you have a cool idea or application... process the CRISPR loci to generate RNA, which guide a Cas nuclease to the specified exogenous genetic...engineering. The diversity of the CRISPR/Cas systems provides powerful lines of defense against invading phages...technology? Let us know.  References Ishino Y, Hideo, S, Makino K, Mitsuko A, Nakata A. J Bacteriol. ...

  3. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    ...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ...structural understanding thereof have the potential to provide improvements to existing technologies based on ...PubMed PMID: 29915061.        A new method to identify proteins occupying a specific genomic locus: Caspex... as an affinity tag. This ‘Caspex’ construct is guided by a pre-defined sgRNA sequence to a specific genomic...preprint posted July 4, 2017 A new method to identify proteins occupying a specific genomic locus: C-BERST... and H2O2, APEX2 labels electron-rich amino acid side chains (e.g. Tyrosine) with biotin. These biotin-labeled...spectrometry allowing a researcher to specifically identify proteins that are enriched in the 20 nm radius...

  4. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...kinetics of the target ligand. Another important consideration is whether the affinity of an experimental protein-ligand...Proteolytic removal of the fusion protein could provide a more relevant kinetic analysis, but the trick...immobilization through biotin-avidin binding could provide a true measurement of high affinity (i.e. sub-nanomolar.... MBP has a similar function as GST in that it provides a method to express a foreign protein in bacteria...biotinylate the lysine residue in the AviTag. As a side note, we found that adding a TEV linker dramatically...that we couldn’t find an all-in-one plasmid that provided an easy way to biotinylate a fusion protein in...biotinylation and purification strategy outlined here provides a basis to determine binding kinetics using a ...

  5. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...transposon? The piggyBac™ transposon was originally identified in moths in 1983, but wasn’t successfully used... that have negative impacts on fitness. One consideration when making a transgenic cell line with the ...the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon system can...donor-plasmid, you could use a single-stranded oligodeoxynucleotide (ssODN) as the donor [6]. The problem with... his specialized knowledge to help researchers identify the right technologies and strategies for their...sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,”...Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted and unwanted target locus modifications...

  6. The MTA at Addgene

    Type
    Blog Post
    ...’ll be confident in what to expect. No system, including MTAs, is perfect, but it does provide consistency...1995: the UBMTA. Because biological materials are widely varied with many properties, having well-understood...Instead, we facilitate the signing of MTAs by the Provider and Recipient (in Addgene parlance, the depositor...plasmids are distributed under the UBMTA, which is widely used across academia. It’s our process that differs...to note that it’s the depositing institute that decides the terms of distribution for their plasmids. ...Webpage, help center articles, and our MTA YouTube video for more information. Happy transferring!    Many...

  7. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    ...This post was contributed by Deborah Sweet, Vice President of Editorial at Cell Press. Almost everyone who...some point. Usually it comes up when a researcher decides on a new project and begins by trying to reproduce... various different versions of checklists and guidelines to help authors understand what they are expected...NIH) now look for data management plans and for evidence of sharing for grant renewal—but there is clearly...hiring, funding, and promotion processes, I’m confident that everyone’s enthusiasm for actually making...Sweet from Cell Press. Deborah Sweet is the Vice President of Editorial at Cell Press.  She trained originally...editorial work to focus on a broader role as Vice President. Her current responsibilities encompass overall...

  8. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...without changing them. Thus, it has been widely adapted for a wide variety of applications: base editing,...fusions localized to specific loci based on the guide RNA sequence. A year later, they developed CRISPRainbow...RFP) to the genomic loci of interest. Pairs of identical hairpins within one gRNA will result in a blue... chromosome-specific loci in the genome and the guides are relatively unstable.   Figure 1: CRISPRainbow...deliver CRISPR-Sirius into cells. They named this guide sgRNA-Sirius-8XMS2 which will recruit the HaloTag...Find plasmids for your research Read our CRISPR guide ...

  9. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ...brilliant ideas on how to solve many therapeutic and pre-clinical problems! Who knows, perhaps the ideas spurred...students play, you can observe, answer questions, and guide them to productive strategies while noting any gaps...their knowledge. Practically speaking the format I decided on was to have two teams of 4 people face each ...with them. My impression was that they had clearer ideas about gene therapy in the following lessons. And...freely share your projects with us! This exchange of ideas will help improve life science and biomedical education...molecular biology educational resources Find protocol videos to teach basic lab techniques Plasmid Land: The...

  10. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    ... took classes in biochemistry, microbiology, epidemiology, and bioanthropology. Studying human health ...doing in the lab and the interactions I was having outside of the “ivory tower.” I’d spend the afternoon lovingly...survived this long without downing any dihydrogen monoxide (H2O). Studying science communication as a PhD...quality of the study design or by their preconceived ideas about the controversial medical issues. Participants...beliefs even though all three texts were otherwise identical. Likewise, when participants read an abstract ...in a behaviour called the “biased evaluation of evidence,” in which people (conscious or unconsciously)..., looking for reasons to dismiss them. There’s evidence to show that people with greater levels of science...

  11. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    ...yoga classes. Get outdoors. Taking a walk or a bike ride is a great activity for both exercise and clearing... times of stress due to #COVIDー19. Receive and provide support while being safe. @GHP_HarvardChan — Shekhar... (individual development plan) from Science to identify skills to work on for the next steps in your career...networking events. Download Addgene’s Science Career guide to learn more about career paths and things you ... Everyone. Plan out new experiments carefully. Consider submitting a registered report, where peer review...is based on the methods, not the results. Also consider using this framework to design your future experiments...Technique. Co-work virtually. With co-working, the idea is not necessarily to chat, but to mimic your usual...

  12. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    ...research from bench to bedside is, in addition to thorough ethical considerations, in making sure that ... our CRISPR guide Find CRISPR plasmids by function Check out our genome engineering guide ...scientists and the public together to build consensus guidelines, but there was never a specific timeline for ...shuffling our feet on necessary actions to review and decide on the future of genome editing technology. We ...Harley and Lexi, and a clutter ofoutdoors-only spiders, all of whom are named Charlotte, all of whom are...

  13. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ... AAV (rAAV) vector (Figure 1). Everything else outside the ITRs gets left behind.    Figure 1: Recombinant....com. ITR structure  ITRs are symmetrical 145-nucleotide sequences that flank the ends of the single stranded...Figure 2). Additionally, the ITR contains a 4-nucleotide Rep binding element (RBE) that serves as a binding...sequences. GC tails are often located immediately outside of the ITRs in the plasmid backbone and do not ...since analysis of individual reads allows for identification of non-dominant ITR sequences.  Restriction...Capsids More resource on Addgene.org Addgene's AVV guide AAV Viral Preps References  Earley, L. F., Conatser...Alexander, I. E., & Logan, G. J. (2019). A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated...

  14. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ...Services - includes videos on general resume writing, as well as additional videos with emphasis for graduate...Relations Director for Curiosity Collider (hyperlink https://curiositycollider.org/) , which she co-founded, ...applying for the position?   Here are a few things to consider when you're preparing your resume for a non-academic...you only have two pages, you need to have a good idea of what should be highlighted on your resume, and...postings and look at the jobs you are interested in. Consider listing the skills you already have, and the skills...

  15. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...terminal repeats; U3-R-U5 regions found on either side of a retroviral provirus (see below). Cloning capacity...Protocols: General Transfection The Addgene Retrovirus Guide  ...

  16. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...collection that grows on a daily basis, the lists below provide a snapshot of relevant resources and a good jumping...the gene you’re looking for on this list, please consider searching directly on our website. In-exhaustive... from a Set of Mitotic Chromosomes. Samwer M, Schneider MWG, Hoefler R, Schmalhorst PS, Jude JG, Zuber...DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies bromodomain protein ZMYND8 in chromatin recognition...receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5)...chromatin and the epigenome. If you have been considering some new techniques, the posts below may steer...

  17. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...can be engineered to be cell type specific. Once inside the host cell, the virus unleashes its cargo – ...transduction efficiency and can effectively infect a wide range of cell types that are typically ‘difficult...viruses have a low immunogenicity which makes them ideal for applications with whole organisms. Finally, ...right approach for your experiment, you’ll need to decide exactly which type of virus to use. Types of viruses...genome into the host genome, setting up permanent residence. They primarily infect dividing cells, as their...signals which can traffic their viral particles inside the nucleus, without the need for envelope disassembly...clinical applications, viruses are true bench to bedside tools. Keep an eye out for our upcoming blogs on...

  18. Hot Plasmids - May 2022

    Type
    Blog Post
    ...subcellular localization peptide. By combining three different localization peptides with four different fluorescent...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ...nanobody-GFP interaction). The modularity of the system provides researchers better flexibility and makes the approach...therapeutics to the CNS, and with reduced systemic side effects, by allowing the user to select for variants... those shown here, only AAV.CAP-B10 causes brain wide expression, with no liver expression, after retro-orbital...mixed-subclone system using mStrawberry, localization peptides, and reporters that drive the expression of the...

  19. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    ...the lab and you're sweating the small stuff? Identifying the Issue You can’t fix an experiment by simply... work. Draw your plate maps onto your plates, alongside your physical copy. If you’ve got messy handwriting...experiment than it is to rerun an experiment, so consider your prep an investment in accuracy.  Bench setup...you add them. Moving your reagents from your left side to your right as you use them can help you remember...to whom I am forever grateful, I started setting aside 3-5 minutes to sit quietly at my bench before the... to a protocol if you’re in a hurry, but let’s consider it this way: When you’re stressed or rushing, ...20Blotting.pdf  More resources on Addgene’s blog A Guide to Getting Started in Undergrad Research Early Career...

  20. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ...Encourage your close peers to come along for the ride, and consider applying to several opportunities. In an ...academic resources, we created STEAMid, for a nationwide database of  STEAM internships for high school...order to gain a competitive advantage this cycle, consider the advice below:  1. Eligibility check Scrutinize...and grades that serve as automated filters of consideration. If you are worried about any eligibility provision...to writing a solid personal statement will pay dividends. This is the only opportunity where you get to...ahead for success. Every student should use these guidelines consistently across a number of years as you ...

  21. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...Microscopy, University of Amsterdam. Before you decide which car you want to buy, it is worthwhile to ...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g... the protein of interest by a 2A self-cleaving peptide (Goedhart et al, 2010). An explanation of the assay...proteins (with the same reference protein) under identical conditions, a ranking of the practical brightness...practical brightness. Therefore, practical brightness provides a better picture of what can be expected in a ...theoretical brightness. Consequently, it makes sense to identify a couple of promising fluorescent proteins and...mimicking the future application. In figure 2 we provide an example of the comparison of the practical brightness...

  22. Addgene’s Viral Service - Why Virus? Why Now?

    Type
    Blog Post
    ...Viral Service. With their support we are able to provide faster access to a larger number of viral preps...Karpova’s lab from Janelia Research Campus have provided valuable insight into AAV production. Finally...advice and encouragement. Why virus? Why did we decide to take on this project? Addgene’s mission is to...plasmids in a ready-to-use viral format. Having identified this need in the community, we approached it ...control, generates  educational materials, and provides viral vector technical service (details about ...expertise in answering questions on virus and helping provide educational resources on this collection. With ... we could ship internationally and be ready to provide extensive logistics support for export and shipping...

  23. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...modification of multiple genes is often necessary. Guide RNAs (gRNAs) are commonly packaged in 400-500 bp...terminator. These relatively large cassettes (considering the gRNA itself is ~100 bases) limit the number...cleaved precisely as predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified ...tRNA-gRNA-tRNA constructs. gRNAs with any 5’ leading nucleotide could be processed, a distinct advantage over...over other methods that favor certain nucleotides at the 5’ end. PTG construct schematics. Scissors indicate...frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct loci. They also examined...this publication at Addgene Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing...

  24. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...transformed the palette of experiments possible in a wide range of organisms and cell lines. In C. elegans... on repair of a conditional-lethal mutation to identify edits with minimal screening effort. Selecting...504 restriction digests of PCR products used to identify a single 2xFLAG knock-in by direct screening. ...several aspects of editing. First, they devised guidelines for designing single-stranded repair templates...from a single injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding...non-homologous sequence or a series of single-nucleotide substitutions along a 1.5 kb tract, all without...CRISPR in C. elegans The Meyer and Mello labs also identified several experimental set ups that could improve...

  25. Addgene: The First Twenty Years

    Type
    Blog Post
    ...across the globe.  “Read the Addgene guide: https://www.addgene.org/guides/crispr/” — SirHamhands, responding... scientists. She, Kenneth Fan, and Benjie Chen decided to start an open science nonprofit that would facilitate...navigate. This meant we could grow accessibility alongside our repository.  “I’m sure you hear it all the...created are for the community. More than 100 labs worldwide use our plasmids. I hope they contribute to great...we realized we could use the same principles to provide recombinant antibodies, another plasmid-based product...including our new Antibodies 101 eBook, the Antibody Guide, and two antibody animations.    A blast ...

  26. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...is planning or might need to plan for a career outside academia (that would be pretty much all scientists...usually visible to you (unless they’ve chosen to hide it). While most people use LinkedIn messaging, not... should be asking yourself if it’s really a good idea.  Pro tip: Don’t hit the connect button unless you...your network. If someone has connected with me, I consider them fair game for connecting to others.  Pro ...can type a full note with good format without accidentally sending before you’ve had a chance to proof ...including and especially the hiring manager. This is considered intrusive and it’s still connecting with a stranger... people’s news and updates. Share your own good ideas, events and favorite resources. Respect the network...

  27. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...sequencing. Sequencing the 16S rRNA reveals the identity of the organisms present while sequencing of all...gives clues into what the microbes are doing. But aside from sequencing, scientists can also study the microbiome...limited and many microbes are difficult to culture outside of their natural environment. Thus, the Harris ...tested this system in vitro and in vivo. They identified cells from 19 genera across four phyla that have...conjugation in vivo studies. However, they only identified seven genera in in vitro experiments, possibly...Clostridium difficile. As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect...Monitoring microbial population dynamics in the gut can provide insight into how the microbiome responds to infection...

  28. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ... manuscript whose whole purpose is to provide detailed evidence of knowledge new to mankind. And you probably...logical to a layperson. When you’re communicating ideas that you think are clearly connected, step back ...overwhelming your audience - like the professor who once decided that to teach high school students PCR, they first...links to pay attention to. If you’re watching a video, you now have moving images, noises and music, spoken...olds is unlikely to be able to follow a YouTube video on PCR, while a senior lab scientist is likely to...distraction, such as pausing or turning off the slide deck (or reminding them to put their phones away...

  29. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...in cells? This question is largely unanswered. Ideally, fluorescent proteins should act as inert modules...understand how well fluorescent proteins approach this ideal situation, cellular assays have been proposed. Early...homodimerization tendency. This assay employs a peptide derived from cytochrome p450, CytERM, to direct... looking cells can be counted. This assay will provide information on the dimerization tendency in living... interest will be problem-free. In figure 1 we provide an example of the OSER assay with a number of fluorescent...mScarlet-I shows correct ER labeling, which is good evidence that mScarlet-I acts as a monomer in the cell ...preprint, or documenting it in some other way. Besides the data, new plasmids and/or cells or organisms...

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

    Type
    Blog Post
    ...LC3 doesn’t provide information about the contents of the autophagosome nor does it provide information...an autophagy inhibitor. While this method does provide a quantitative measure of autophagy, it only measures...studying autophagy. This method relies on the identification of autophagic structures based on morphology...morphology. Autophagosomes are relatively easy to identify: double-membraned structures containing undigested...with the vacuole or a lysosome are trickier to identify because their contents can be at various stages...Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, another fluorescent pH-biosensor...
Showing: 541 - 570 of 1104 results