...plate. You can use this plate to start liquid cultures. Lastly, you can start small overnight liquid cultures...detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase) using...PCR reactions is nearly identical to preparing a standard PCR reaction: combine template, primers, polymerase...polymerase, and dNTPs and then incubate with a standard PCR thermocycling program. One key difference is ...leftover bacteria-water suspension and use them to start cultures of your positive clones. If you want to...the initial heating step of the PCR reaction. A standard Taq polymerase is sufficient. Controls: Controls...Make sure to use an appropriate molecular weight standard for reference and to add a loading dye with glycerol...

...regulations, differing customs processes, and the distances the packages needed to travel meant the accessibility...using couriers that refill dry ice if the package stays in transit for a certain number of days and tracking...Addgene distributors Distributor Partnership Start Date Country Served Beijing Zhongyuan 2012...information about the materials in our repository. We started to increase accessibility to our repository by ...types, and more, all of which help a researcher understand if a material is right for their experiment.   ...introduced guides — practical primers to help users get started with tools like CRISPR, lentivirus, and optogenetics...darker blue indicates more views from that country. Stats and figures are helpful, but our favorite measure...

...and some rising stars in the world of microbiology, some before they had even established their own labs...Tweet - The conference hashtag I had, however, started a Twitter account, and was following the meeting...Favorable conditions - Expansion of networks The last stage is Exponential Growth, named after the growth phase...variability was organic, as some people need less assistance than others, but we did have some participants...worth addressing. Clear expectations have been established for future participants regarding the activities...mentoring. For example, Goldschmidt conference, Coastal & Estuarine Research Federation (CERF) Biennial... and microbiology for the last two decades as a staff scientist, but before going to graduate school, ...

...unintentionally remove connecting pathways between other distant regions. They also don’t give any clues about the...Anxiety tests Rodents have a natural preference to stay in dark areas (protected) as opposed to brightly...mouse can move freely on a 4-arm cross and choose to stay on an open arm (risky), or a closed arm (safe). ...On the other hand, the control group preferred staying in the dark compartment (Howerton at al. 2014)....floating on the water surface. The latter reflects a state of behavioral despair, and the more time the animal...genetic manipulations with the forced swim test understanding the genetic background of depression.  One such...memory. We’ll cover these in a  future blog post, so stay tuned!  References Alpár, Alán, et al. "Hypothalamic...

...human genes have a counterpart in zebrafish. Understanding the function of these genes in fish will help...functions for genes, and lays the groundwork for understanding the gene networks that regulate processes from...mutagenesis screens provided a major toehold into understanding how genes control biology (a fact celebrated...for generating transgenic zebrafish are well established (Felker and Mosimann, 2016). One common and robust...zygote transforms into an organism with a clearly established body plan including head, eyes, and tail. Compared...different methods have been developed, relying on established reagents such as CRISPR/Cas9, Tol2, and the high-quality...profiled 38,731 cells from 694 embryos over multiple stages and reconstructed the patterns of gene activity...

... for numerous non-academic careers. Still, many start their scientific careers hoping to follow the traditional...Possible roadblocks only become obvious at rather late stages (i.e. postdoc level) and can include the pressure...candidates are highly experienced but often have to start pursuing other options. While some non-academic ...existed, but did not consider them for myself. I started picturing myself in a technical support role only...possibly this same reason, it can be one of the more constant and safe corners in a company, less exposed to...where someone tries to “optimize” a particular standard lab protocol. For example, someone may attempt...then complain about high background. In many instances, variations of the product’s specific application...

...Background on neuronal tracing A key aspect to understanding the brain’s function is knowing its architecture...importantly, any connections the neurons make to other distant brain areas. It is desirable for tracing compounds...the direction of information flow in order to understand which brain regions communicate with each other...decades was essential to build up our current understanding of neuroanatomy and neural connectivity, providing...advantage of AAV vectors is that there are well-established protocols for their production and purification...expressed a fluorescent marker in areas close or distal to the injection site across the entire brain, ...connectome”, to be visualized in a uniform and standardized way (Oh et al., 2014). Images yielded from this...

...scientific titles generated from the United States Standard Occupational Classification System  and Wikipedia... in the United States (0.428). 4. The #1 (re)tweeted domain by scientists was Instagram Social media and...they are based on the US Occupational Employment Statistics (OES), but international Twitter users were included...don’t use Twitter, are you interested in getting started? Reply in the comments below or tweet us your reply.... Other Resources Check out the Top 50 Science Stars on Twitter Want suggestions on how to use social...

...Rab11, and Rab35 to understand their roles in forming the cilia that help establish left-right organization... and limitations – for example, microtubule-destabilizing drugs are impossible to target to specific subcellular...available as lentiviral plasmids if you want to make a stable cell line. This design allows highly localized,...reversible microtubule severing and has very low dark-state activity and toxicity. All of that means this light-powered...dissect the roles of three Rab proteins during a key stage of zebrafish development (Aljiboury et al., 2023...After 30 minutes of blue light illumination, substantial PA accumulates in the plasma membranes, indicated...moderately-enhanced 10x and 30x SuperPLDs avoid dark-state activity and cytotoxicity better than the 50x and...

...suspenseful. 4. Start preparing your talk early - like today Don’t wait until you need to speak to start creating...’s attention will start to wander when you are supposed to be done. They will start looking at their phones...future boss in the room, so it is a good idea to start thinking about public speaking early and often. ... audience is that it can help you avoid making mistakes. I once made a joke about social science to an...Describe how you pushed to a solution or overcame obstacles. Take time to consider the order of your narrative...t be hard on yourself if you stumble or make a mistake. Move on, the audience doesn’t really care about...

...how CRISPR can take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and...ART-induced viral control found in most HIV-1 patients. Obstacles for CRISPR HIV-1 therapy Both of these methods...survive, but preventing re-infection would require sustained expression of Cas9/gRNAs, which could result in...enough T-cells could be reached to ablate a well-established viral reservoir. A second challenge for CRISPR...binding. It’s also possible that HIV-1 may evolve resistance to CRISPR therapies through PAM or seed sequence...multiple drugs to lower the odds of developing resistance. Shortly after Kaminski et al. published their...not ablate viral function, but rather promote resistance to CRISPR/Cas9. While these results are discouraging...

...to put together DNA fragments. As a fun way to start, I highly recommend watching this entertaining video...vector of choice as shown in the figure below. To start, you need to have DNA fragments with regions of ...not work well if the ends of the fragments have stable single stranded DNA secondary structure, such as...Gibson assembly isn’t ideal for synthetic biology standards, which rely up on the re-use of parts between ...One way around this is to use a combination of standard sequences with overlap regions, such as in the...to cleave the 22kb vector. When followed by the standard Gibson assembly techniques described above, this... to get mixed up with Gibson assembly (they all start with a G!) but actually work by significantly different...

...antibiotic resistance cassette and you aren’t introducing an additional antibiotic resistance cassette ...where the inserted DNA includes an antibiotic resistance marker. After transformation, plate the cells...into the plasmid backbone contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing...cells carrying plasmids that contain a kanamycin resistance cassette will grow. However, often times, the...often used for cloning in yeast since antibiotic resistance spontaneously arises in yeast. For example, Markus...occurring genes for cloning purposes (antibiotic resistance genes, toxin genes, and the sacB gene). By tapping...

...that efficiencies of stable gene transfer using SB100x were comparable to stable transduction efficiencies...enzyme.2.) The transposase enzyme is expressed (red stars) and binds the inverted repeats (green); and an ...determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature...This new tool kit allows the rapid generation of stable transgenic cell lines which can robustly express...Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol J. 2015 Feb 4...hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun...

...don't, of trying to understand your completely unexpected findings, or understanding why your experiment...nothing if not solving your way through a series of obstacles in pursuit of achieving knowledge. Or, as we in...result of competing desires or (3) the presence of obstacles that need to be overcome. Conflict, and its inherent...them want to know more. This could be by outright stating the big-picture problem, by asking your audience...provide enough information so that the audience can understand the conflict. A science-based conflict gives ... and they all can be effective in the right circumstances.   However, in the midst of all these storytelling...

...bacteria have just reached stationary phase, but the cells have not started to die off. Have any questions..., 2020. Now that we know all about antibiotic resistance genes, let’s consider another basic element of...cell. Figure 1: A plasmid map showing the standard features of a plasmid. So, how do you choose...researchers can ask themselves a few questions to get started: Will the plasmid be used exclusively in E. coli...will compete for the same machinery, creating an unstable and unpredictable environment. As a rule, plasmids... subculture directly from glycerol stocks, agar stabs, or liquid cultures. Freshly transformed cells also...

...alternative” anymore.   Since most scientists start out in academia, it is especially important for ...article well worth reading on this topic, the authors state, “Both explicit and implicit aspects of today’s ...”.   In fact, there are a lot of good reasons to stay and good reasons to go.  Here are some questions...calendar Academic researchers must be extreme self-starters and especially good at self-promoting—No one is... is going to tell you to pick up the pace Constant worry about grants and renewals What is different ...about something like this without sufficient data? Starting to learn about the diverse options and how to ...Science Careers How to Negotiate a Successful Lab Start Up Budget Read Our Management for Scientists Series...

...Scintillon Institute, Allele Biotechnology, Florida State University, Indiana University, and the Karolinska...protein wavelengths, it can be imaged either with standard green fluorescent protein filters or yellow fluorescent...brightest teal fluorescent proteins that were photostable and had a high 480/530 nm emission ratio. They... in signal relative to mCerulean imaged with a standard CFP set. mTFP1 can be used as a FRET donor to ...CFP484. Collaborating with scientists from Florida State University, they used a mutagenesis approach and...mWasabi is 1.6-fold brighter than EGFP and is photostable. It also has narrow excitation and emission peaks...Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein...

...collaboration for gene classification This inquiry started when Neil Thompson, a research scientist at MIT...at the time, got involved. Along with Philipp Pfingstag, a visiting graduate student from TU München at...was moved exactly as is from one organism to a distantly related organism, the gene may not express well...not rare codons and many genes where there are instances of rare codons, sometimes intentionally modulating...modulating expression in ways we don’t quite understand yet,” Kunjapur says. Percent sequence identity was...showed a growing trend of sourcing genes from distantly related organisms. In 2006, less than 1% of the...  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows ...

... and conditions to understand exactly what your antibody recognizes.  Understanding how the available ...the protein folded, changing it from its native state to a less folded, or denatured, one. This enables... protein of interest while it’s in a transition state or specific configuration. So how do you ensure ...protein as it exists in your assay?  First, try to understand, to the best of your ability, what factors may...during your sample prep process, imagining how it starts and if any steps could affect its configuration...assay that uses similar sample preparation - for instance, if you cannot find an antibody validated for ...