...even come [to grad school]!” He asked if she had considered other careers, which she hadn’t.  He then said... didn’t know if she liked science writing. She decided to start a blog to test it out and discovered she...communications planning.” To beef up her resume, she decided to complete a certificate in Strategic Communications...these difficult pieces: “It’s probably the best.” Besides working through the tough assignments, the skills...important for her work include: “Listening. Evaluating evidence. Critical thinking. Being able to learn things...

...to $47,476 per year. This post discusses how a nationwide injunction against the FLSA is affecting universities...injunction against the FLSA updates was granted nationwide. Again, we at Future of Research believe that...higher than the level being raised to now - across a wide consensus of academic stakeholders. However, while...collecting data on the effects of the injunction nationwide and, at exactly one month following the injunction...plans, but have since reversed their position and decided to allow individual departments to go ahead with...

...time-consuming aspects of traditional cloning is the identification of clones that actually contain your insert...enzyme-mediated disruption of ccdB, leading to positive identification of desired clones.   ...Becomes a powerful... insert of interest. Hence correct clones are identified much more efficiently, as those that do not contain...ultimately perform the same function, there is some evidence that ccdB Survival™ can be more difficult to transform...E. coli do not contain the F plasmid (and are considered F-); however, there are a few popular lab strains...

...blogger Didem Goz Ayturk with edits and updates from Addgenies Karen Guerin and Susanna Stroik. Didem Goz ...administered in an organism. These features make AAV ideal for human therapies and in vivo laboratory experiments...therefore the transduced gene is episomal and does not divide and replicate in the same way the host DNA does...Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All Viral Vector Resources...

...thousands of plasmids, using both mass production genome wide screens and the more traditional “one at a time”...time. We also handle the MTA, QC the plasmids, provide helpful resources, and, on the rare occasion things...organized lab saves time and money across the board. Consider alternative cloning strategies There are many ...either don’t scale or have limited scaling. The downside of high-throughput work is that you give up a ...engagement with and use of repositories like Addgene provides large-scale savings. How large? Using Doench’s...

...affects protein function (Bornhorst 2000).  One downside of polyhistidine tag-based purification, however...contamination, such as mammalian production systems, consider using these more selective substrates along with... containing a polyhistidine tag, you’ll need to decide whether to add the tag to the N- or C-terminus ...the opposite end.  If neither placement works, consider purifying under denaturing conditions. Unlike ...References  Resources on addgene.org Addgene Antibody Guide Ready-to-use antibodies from Addgene More Resources...

...postdoc offices or professional organizations can provide referrals. There are bad coaches out there, so ...the one for the job. A professional coach will provide an initial consultation meeting or call for free...here are a number I have come across for you to consider. Feel free to add other suggestions in the comments...Scientist. Their motto is “We Turn Academics into Confident and Successful Industry Professionals”. This larger... 6 Steps to Submitting a Resume That Gets Seen  Guide Your PhD with An Individual Development Plan Learn...

...across the university. Collective bargaining may provide a better deal for students in areas such as teaching...philosophical landscape of the scientific enterprise will provide challenges and opportunities for us all.  For example...efforts to better account for and track postdocs nationwide. On this level, FoR is currently compiling a ...further increases will be challenging, but will be aided by the development and implementation of new strategies...create tools to better understand human disease.  Outside the lab, he is interested in communicating science...

...transduction. Once you’ve harvested, it’s generally a good idea to titer your virus. Getting an accurate count of... blog and be sure to check out our viral vector guide pages to get up-to-date tips for your virus experiments...10325307. PubMed Central PMCID: PMC84922. 5. Ugai, Hideyo, et al. "Stability of a recombinant adenoviral ...immunotherapy." Acute Leukemias VIII. Springer Berlin Heidelberg, 2001. 291-306. PubMed PMID: 10634178. 8. Murnane... on the Addgene.org Check out Our Viral Vector Guide Pages Find Virus Protocols Order Virus ...

...-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...Hemizygous mice can be included in breeding strategies identical to those used for breeding heterozygous mice. ...strains that have been crossed, there are many considerations for effectively breeding your mice. Because...that express Cre but not loxP. With this two-part guide to mouse modeling now complete, you should be prepared...

... two approaches: single-cell RNA-sequencing to identify neuronal class and deep sequencing of neurons ...database specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures...imaging processing. The combination of these methods provides unique opportunities to develop computational ...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...of CRISPR targeting vectors that each contains a guide RNA and repair template of your choosing (Schwartz...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... than the more efficient single-stranded oligodeoxynucleotide donors (ssODN). So how can you use CRISPR...Resources page.  CasPhi is 70 kDa Cas protein identified exclusively in huge bacteriophage genomes. It...dual-targeting hybrid gRNA library can be used to identify genetic interactions between paralogs. The exon-deletion... viral aliquots from our plasmid collection to provide ready-to-use viral preps. Here are some of the ...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was applied...photostable than fluorescent proteins, making them ideal for single-molecule fluorescence techniques. After...measured pixel and the soma. The initiation site identified in panel (C) is indicated; red line is exponential...research! If they do (or if they don't!), please consider sharing your experience with other researchers...

...maintain balance – encouraging cells to grow and divide only at the right times and in the right places...trying, as the circulatory and lymph systems are not ideal environments for many cell types. From proteins ... in the development of different cancers and identifying the roles of key proteins involved, with the ...Resources page, investigators may select a pathway and identify any genes within that pathway that are of interest...References 1. Martz, Colin A., et al. "Systematic identification of signaling pathways with potential to confer...

...multi-view imaging can provide enhanced resolution (8,9).  Thus, LSFM is ideal for 3D reconstruction of...couple-charged device (EMCCD) or complementary metal-oxide semiconductor (CMOS) sensors, it takes only a few...z-stack (8).  One limitation of LSFMs using single side illumination, especially with large tissue, is that...and acquire images of a sample from two opposing sides and merge the images, such as in the Ultramicroscope...

...thus tracking the localization of that biomolecule inside living cells. FPs classified by the emission color...range, there are other traits that need to be considered when choosing an FP: Unique categories of fluorescent...distance between the two FPs is <10nm, and should be considered when labeling proteins that interact. Indeed,... coefficient and quantum yield of the protein, divided by 1000. In many cases the brightness is compared...greenfluorescentblog.wordpress.com.   More Helpful Websites & Resources: FP guide at Addgene Interactive Visualization of Fluorescent...

... degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation... can be assembled. For example, because CpG dinucleotides are underrepresented in most vertebrate genomes...and MluI recognition sites, which contain CpG dinucleotides in their 6-mer sequences, are rarely found in...for cloning genes from organisms with high CpG dinucleotide representation. The specific restriction enzymes...

...Heinrich, W. (1983). Worldwide epidemiology of human adenovirus infections. Am J Epidemiol, 117(4), 455–466...length. Human adenovirus type 5 (hAdV-C5) can be considered the prototype, although many viruses infecting...adenoviruses zoonotic? A systematic review of the evidence. Emerg Microbes Infect, 8(1), 1679–1687. https...

...After learning of it's continued popularity, I decided this protocol would be a worthy first contribution...protocols. After my introduction to protocols.io, I decided to research what other online (and mobile) tools...www.addgene.org/ If you’re working with plasmids and you decide to share them using Addgene’s repository, you can...Protocols OpenWetWare.org (OWW) is a wiki that provides a community for biologists and bioengineers to...and allows scientists to contribute protocols or provide links to protocols on their own websites. We're...

... but many find it helpful to obtain support and ideas from those who seem to be managing. Being reminded...balance tips, we need to look to for support and ideas to manage our stress. If you are saying to yourself...appreciate this – and they will be happy too. On the flip side, if you are not happy at work, you probably need... you will get help when you need it. On the work side, join a professional community that you can count...an emergency and this will make you much more confident about their safety when they leave home which ...

...ternary complex with the multiple myeloma drug mezigdomide and ubiquitination substrate Ikaros (IKZF1ZF2...important functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024...CRBNmidi as a promising tool for future structure-guided drug design and studies of protein ubiquitination...showed greater photostability and also matured considerably faster, with expression levels comparable to...expression and removing the sgRNA plasmid after identifying successful knockouts limits further cutting and...

... you have these rules (word limit, formatting guidelines, etc.) you have some criteria to shape the document...research papers that I needed to read. Once I had an idea of the themes in the field, I searched for recent...field. I made sure to find information from genome-wide studies, as well as results from smaller and more... or popular papers, but looked for papers from a wide range of authors. Just start writing When I first...Plasmids Review the CRISPR Field with Our CRISPR Guide Pages ...

...Hedgehog but this is not a direct reference to the video game character but rather to an extremely important...Christine Nusslein-Volhard and Eric Wiechaus first identified the hedgehog gene in Drosophila. They observed...nonsyndromic holoprosencephaly where the brain fails to divide properly into the right and left hemisphere. Addgene...origins of the name but some of our Addgenies had an idea: sqh mutant fly ovaries had “squiggly” actin fibers...Additional resources on the Addgene blog Read our Quick Guide to Working with Drosophila Read our Plasmids 101...

...negative staining. Empty vector particles can be identified after negative staining and appear darker than...been used to titrate AAV, but this method is not widely used today (Fagone et al., 2012). PCR-based methods...from different sources in the same experiment, consider retitering both AAV in your lab. The absolute ...does not give an indication of infectious titer, consider titrating each lot of AAV in vivo to determine...Additional Resources on the Addgene Blog Important Considerations When Using AAVs Addgene's AAV Quality Control...

... one bacterium sharing genes with another, an accidental transmission of genes from one bacterium to another...tried and true process of evolution. Watch this video showing bacteria evolve antibiotic resistance on...different reasons. Exposure to antibacterial drugs provides a selection pressure, killing off bacteria that...prescribed to them This may seem counter-intuitive considering the other three scenarios, but a short treatment... antibiotics and which ones aren't?     This video shows how to perform a Kirby-Bauer disk diffusion...

...Mortensen et al., 2019). Our method might provide some useful guidelines for the development of transient transformation...transformation systems for other species as well. Things to consider in developing a transient leaf transformation ... promoter in Agrobacterium. This can be easily avoided by the use of introns, which will be removed from... to quickly assemble vectors and test different ideas. We are a huge fan of the Golden Gate based MoClo...

...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...that evolved in the last 10,000 years (which is considered recent). They have diverse traits with good genetic...control who a female wants to mate with and if she decides to raise their offspring or leave them to their...a cool fluorescent indicator system to rapidly identify recombinants. Arthropods (Jason Rasgon) This lab...

...Sequencing results are analyzed to confirm the identity and integrity of the viral genome and the absence...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...negative staining. Empty vector particles can be identified after negative staining and appear darker than... monitored throughout production, ensuring the identity of each sample from the DNA tube all the way to... preparation. After production, we confirm the identity of the transfer plasmid in the viral preparation...

...post on viral vector parts. Promoter specificity Aside from choosing a promoter based on type of RNA transcript...transcript     Description Expression Additional considerations CMV General expression mRNA Strong mammalian... phosphoglycerate kinase gene.  Constitutive  Widespread expression, but may vary by cell type. Tends ... used for Description Expression Additional considerations T7 in vitro transcription/ general expression...therapeutic uses, and in those cases it's important to identify the right tissue-specific promoters as described...

...Addgene's Viral Vectors Guide Addgene's Chemogenetics Guide Addgene's Optogenetics Guide  ... our Viral Vectors 101 eBook!  This practical guide to using viral vectors in the lab is a helpful resource...

...so we’ve been busy updating our CRISPR guide. Our CRISPR guide is a comprehensive resource that covers...week at #AddgeneCRISPRweek. We updated the CRISPR guide! If you’re not into Twitter, we have some exciting...for planning your CRISPR experiment. For a quick guide to CRISPR, download our CRISPR Cheat Sheet....

... Summers, 1998). Multimeric plasmids are also considered to be less stable and more prone to being lost...other methods below should help in multimer identification.   Fig. 2: Gel image of Plasmid 61564...used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services using long-read...within a plasmid and, therefore, allow for the identification of plasmid multimers. Capillary Gel Electrophoresis...laser-induced fluorescence (CGE-LIF) for detection and can provide data on the quantity of various topologies and ...

...insights into the process of getting an MD PhD, the inside scoop on developing new CRISPR tools, and the potential... : Alex introduces himself and explains why he decided to do an MD PhD - it's all in the family. 3:56 ...Biologically Inspired Engineering and why Alex decided to work there. 13:22 - 22:07 : Alex's projects ... Resources on Addgene.org Check Out Our CRISPR Guide Pages Find Ready-to-Use CRISPR Lentivirus Learn ...

...this concept in his microbiology lab course, he decided to add a bit of fun. He mixed together 10 E. coli...host strains affect transformation efficiency.  Aside from the Streak Plate Challenge, Westenberg’s course...plasmids here, and check out our streak plate protocol video on Youtube:     Additional resources on the...plasmids at Addgene Check out additional written and video protocols ...

... In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...compatible with the introduction of unique molecular identifiers, which can improve the statistical power of in... of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses...Context-Dependent Germline Knockout in Immune Cells Genome-Wide Screening Using CRISPR Cancer and the Immune System...

...some good guidelines to follow in order to ensure that people can quickly and easily identify what your...the insert. pBACKBONE-Flag-hGene-EGFP Other considerations  If your insert contains a mutation or modification...generally listed as the amino acid change and not a nucleotide change. The proper way to denote an amino acid...

...journals has risen steadily from 1995 to 2013. Considering citations to all articles, Anurag Acharya and...citations for older publications. (As an interesting aside, Nature recently reported that the most cited work...their own libraries and offering another way to consider and quantify the impact of scientific publications...find ways to capture scholarly work that exists outside of traditional scientific journals. Of course, ...

...puzzle fell into place, as they, taken alongside the insights provided by the studies in Tsien lab, covered...victoria (Figure 1) in molecular detail. On the underside of the “umbrella” of A. victoria there is a ring...1980s, another researcher, Douglas Prasher, got the idea that this new green glowing protein could be used...

...genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus hisashii, BhCas12b that...Microbiology Find the Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics...a screening approach, the Jennifer Doudna lab identified three different Cas12a inhibitors. Some of these...sheet Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited...

...communication? It is important to identify your audience and consider their background. Social media is...contribution can either be an article, a short post, a video, or a piece of sci-art.  What are your top tips ...communication posts Sign up to receive our science career guide Learn about different careers in science communication...

...absence of certain markers on the surface or the inside of your cells. While proteins themselves can emit...fixable dye stains that react with free amines on and inside the cells and are thus typically used for distinguishing...vector.) In our flow experiment, we'll want to identify CD45+ cells (CD45 expression) and assess our transduction...should exclude APC-Cy7 from the rest of our panel. Besides some background cell death constantly happening...Compensation Resources on Addgene.org Addgene's Antibody Guide Ready-to-use antibodies in the Addgene repository...

...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...is used to detect proteins on the surface of or inside whole cells. Antibodies - typically, primary antibodies...staining. This panel of antibodies is used to both identify the cell subsets of interest from among the mixture...This could be cells or tissue slices mounted to a slide, or whole organs. Because the tissue architecture...IMMUNOGENS, VACCINES, AND IMMUNIZATION. In: Immunology Guidebook. Elsevier, pp 17–45. https://doi.org/10.1016/B978...

.... Although my interests are very clear, I had no idea how to translate them into real world careers. I...this new virtual world, I was worried about how my ideas would be translated through Zoom calls and Google...frequent Zoom meetings helped me communicate my ideas. I was able to express my visions for what my Instagram...with Quintin Marcelino who is Addgene’s Science Video Production Specialist and Arthur Guerra who is a...

...Cre - A useful tool for neuroscientists AAVs are widely used by neuroscientists because they efficiently...interest is expressed.  A cell-type specific Cre can provide you with that control in a modular way. For example...only be expressed in neurons. Indeed, this AAV provides you with a means to specifically activate expression... viral vector blog posts for many more tips and ideas on how to use these fantastic tools in your experiments...

...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ... lab life a bit easier. Since the post doesn’t provide an exhaustive list of all the apps out there, we...reasons: 1) I think phage are super cool and have considered getting a tattoo of my favorite molecular biology...to screen for proteins that could be useful for a wide variety of reasons. 3) I would love to see more ...

... but if it is not possible consider a phone or video call. If you do decide to use Skype, make sure to... from other scientists doing interesting work. Consider this to be like any other research project. You...

...traction. One way to consider where the open access (OA) movement might be going is to consider some of the places...Access (YODA) Project, “a collaborative endeavor guided by aspirations to promote open science, ensure ...MOOC), with an explosion of courses and content providers and participation from many well-respected and...

...them remain uncharacterized. This represents a considerable barrier to furthering research and developing... an assortment of genes. Using patient data to identify mutations of unknown importance in cancer Previously... Center patient databases, this research team identified 1,049 perturbations in several genes spanning... 33 cancer types. The types of mutations they identified included gene amplifications and fusions, missense...

...Maureen Hanson’s and Rima Menassa's labs made the accidental discovery that laser treatment can photoconvert... Drosophila and mammalian cells, and it may find wide use in biological research. Photoconvertible fluorescent...great scientific discoveries, this one was made by accident. After exciting GFP-labeled plant cells at 405...variants used by Sattarzadeh et al., as well as a wide variety of other fluorescent proteins for your imaging...

...spoke with Daniel about the interplay of these two ideas he said “science is a form of art to me… The problem...to have increased value and is appreciated by a wider audience for more than just its biological importance...of familiar flowers." This new x-ray technique provides scientist with a new method to explore the inner...organisms without destroying the original, which could provide a new tool to study old and unique samples, such...