... science graduate programs are still doing an inadequate job of supporting science trainee transition ...captured in this just released report “Enhancing Dissemination of Evidence-Based Models for STEM PhD Career...to identify the most effective practices and disseminating career training approaches in a way that can...professional development are intended to be widely disseminated by “train the trainer” programs.  Please join...

... a set of techniques that are used to insert recombinant DNA from prokaryotes or eukaryotes into a synthetic...(genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification ...scientists an indication of purity versus protein contaminants. The 260 and 230 nm ratios should fall ~2 - ... being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely endotoxin...

... This blog was originally published on BitesizeBio here. As part of my job ensuring plasmid quality at...sequence Anything less and you might suspect contamination in your sample or consider asking your sequencing...but well worth it. 6. Edit your DNA sequence Finally, when you do see a miscalled peak, don’t be shy... Acknowledgements: Thank you BitesizeBio for originally publishing this and allowing us to share it with...

...plasmids  Plant base editing with a Cas9-adenosine deaminase fusion Caixia Gao’s lab has developed a new plant...plant base editor from an evolved tRNA adenosine deaminase that is fused to a nicking Cas9. This base editor...it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular ...an alternative to the conventional Cre or Flp recombinases used to excise markers. It has the advantage...

...with a confocal microscope, yet I was once again fascinated by the world that reveals itself when looking... both interesting and approachable!   The grand finale of visual impressions was delivered by the ASM ...ASM Agar Art Winners! It is absolutely fascinating to see the living art designed by creative minds with...steady hand. By connecting art and science, a fascinating interface is created that produces loads of inspiration...

...Darrel does 20% clinical work 80% research and was originally focused on becoming an MD but later got obsessed... therapy, and tissue engineering. How the multidisciplinary nature of the Center for Regenerative Medicine...lungs are difficult, but other tissues like the retina have cell-based therapies underway. Chapter 7: ...Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from...

...promoters, N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant terminators and resistance...percent efficiency, simply stopped working. They finally traced the problem back to the endonuclease. “It...

...with different varieties of tags, both N- and C-terminal. Each set of plasmids uses the same restriction... thorough introduction to pCri, please see the original publication cited below. pCri plasmids are available...al. "The pCri system: a vector collection for recombinant protein expression and purification." PloS one..., Zachary T. Britton, and Anne S. Robinson. "Recombinant protein expression and purification: a comprehensive...

...Homologous recombination is the process by which nearly all domains of life repair genomic damage, specifically...their findings, the authors recommend optimal FP combinations for use in yeast imaging, categorized by specific... of yeast tagging modules with each possible combination of protease site (TEV or PreScission), epitope...protease site)-6xGly linker-(epitope tag)-ADH1 terminator-(selection marker). Additionally, Dr. Formosa...

...repress gene expression. In the dark, the NLS is inaccessible and the complex remains in the cytoplasm. Read...editors with minimized off-target DNA editing The original cytosine base editor, BE3, induces a low frequency...validated CBEs with reduced Cas9-independent deamination. They generated these editors using the BE4max... Fusing epigenetic modifiers such as TET1 with inactive Cas enzymes can alter gene expression via acetylation...

... centers in Boston.  I was relieved because it finally feels to me like the postdocs are taking some steps....  His list is dire:  The current system is unsustainable, hypercompetition is corroding values, we are...awards systems need to be radically overhauled.  Finally, the science publishing system needs to be changed... scientists to start addressing this problem.  Finally, one point made by Dr. Graham Walker, Professor...

... the discussion in markers on a whiteboard. The final image is pictured here…she started from a blank ...labs and the importance of the strong, cross-disciplinary collaboration they shared to the work they did...extra fun) who shared first authorship on their seminal, cooperative paper in Science.  Interestingly, ...forever. He emphasized the potential of this extraordinary breakthrough. In fact, the panelists agreed ...

...lines: ones either suspected to be contaminate or easily contaminated, as well as commonly misidentified...informative. Spoiler alert: HeLa is the most common contaminator line.  For those wanting to access Cellosaurus...profiling, you can use Cellosaurus, and CLASTR, to finally know whether it was an 'h' or a 'b' written on ...

...guide to becoming a successful writer. When I originally talked to my PI about becoming a better writer...aspects to it: videos starring Roberto Kolter, webinars taught by Peggy Cotter, Mary O’Riordan, and Victor...don’ts of that week’s piece of a manuscript. The webinars were my favorite part due to the interaction with... the individual figures and results to make the final product with an interesting introduction and concise...

...sequence through a mechanism called depurination. In depurination, the hydrolysis of the glycosidic bond... DNA stability in solution are pH and DNase contamination.  Extreme pH can denature, break, and even change...gene, there are usually extra steps like heat inactivation, protein degradation, and/or the use of chelating...

...some public speaking and a bit of video work for webinars (get tips for your next science talk). I do illustrations...website quite a bit.” Out of all of these tasks, webinars are Hans’ favorite thing to work on. “It’s one...someone to take over hosting them. Since then, webinars make up a larger part of Hans’ workload. “I am...getting abstracts and plans together for my next 4 webinars—I have them stacked close together for the next...

...still free, and different regions of the spinal cord were illuminated. By looking at groups of neurons that...expressed the LiGluR opsin in different subsets of spinal cord neurons using the Gal4/UAS system. This created...dissection of a behavioural module in the vertebrate spinal cord." Nature 461.7262 (2009): 407. PubMed PMID...

...the AAV genome’s two inverted terminal repeats (ITRs); but when recombinant AAV (rAAV) is made in the lab...generates the different C-terminal regions of the Rep proteins. While the unique N-terminal domain of Rep78 and...

... nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs to the...Sternberg et al., 2014). The nuclease lobes are inactive in the absence of guide RNA, preventing Cas9 from...loaded and the DNA sequence is recognized, it’s finally time to cut! The HNH domain, which cuts the target...flexible; it can rotate about 140 degrees between inactive and active conformations (Bravo et al., 2022; ...on this topic are still being made! Conclusion Finally, we can summarize the process from start to finish...

...particles. A single viral particle is called a virion. Finally, viral vectors, which are used in the lab, are ...contrast to bacteria and eukaryotes, which use a binary replication process whereby one cell copies itself...engineering controls — changes made to the virus that eliminate or reduce the risk of infection — researchers ...genetically modified to either limit or completely eliminate their replicative ability. The remaining particle...

...disturbing the tissue for imaging or collection could contaminate it. I have used it for selecting transformation...promoter with self-cleaving 2A site linkages and a terminator. Image from He, Y., Zhang, T., Sun, H. et al....untransformed seed from the same plant beginning to germinate. (Top right, second panel) Transformed seedlings... Biology Department at the University of North Carolina at Chapel Hill. He has a passion for microscopy...

...the first vaccination and after both vaccinations. For the B.1.351 variant, a single vaccination didn’t ... SARS-CoV-2 variants coincide with increased vaccination efforts. So, how effective are the current vaccines...

...next experiment. Nucleic acid based machines Recombinase-based State Machines Enable Order-dependent Logic...inputs are received by a cell. These so-called recombinase-based state machines could potentially be used...development. Read this post to learn more about recombinase-based state machines and how they function. Adapting...them. CRISPR Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening CombiGEM is a technology that...

...Ultragenyx Pharmaceutical in 2017. As one of the original employees at Dimension Therapeutics, she helped...interested in for a long time. “At an early age, I was fascinated by the impact that medicines can have on human...during graduate school at MIT. “I read some of the seminal papers on cell cycle discovery from Lee Hartwell...delivery. In 2018, he interviewed Addgene co-founder Melina Fan and former Associate Director of Viral Vectors...

...preparation approaches described previously require a preliminary double stranding step. The double stranding step... library preparation procedures. This finding eliminates the double-stranding step and reduces both the...confirm the identity of AAV genomes and detect contaminating AAV DNA. In addition, we wrote an open-source...uses the VGS data to determine the percent of recombination present in Cre-dependent vectors and a script...

...sequences on a daily basis, many found these tools inadequate for planning, visualizing, and documenting their...elements such as promoters and transcription terminators, the boundaries are less obvious. We found that.... In an effort to be rigorous, we dug into the original literature, some of it decades old, to provide...mismatches, our algorithm could annotate a feature inaccurately. The best example is fluorescent proteins, which...

...electrical activity through imaging. Since it’s illuminated by red/orange light, Archon1 can also be used...Chimeric channels for neuronal manipulation Recombinases Plasmid Serotype Name 50363  AAV5  AAV ...labeled neurons because background fluorescence from inactive neurons is reduced. 105322  AAV1  pGP-AAV-...densely-labeled neurons because background fluorescence from inactive neurons is reduced.  Additional resources...

.... We will continue to provide you with more fascinating, informative, and actionable content in the coming...genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions, deletions...Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and tired of having...you’ve made the right one. In this post, Addgenie Melina Fan guides us through the basics of the validation...

... embryonic stem cells, they found very high recombination percentages (78-79% after two rounds of transduction...application of iTOP Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to see if iTOP ...events. For these reasons, iTOP is best suited to binary systems in which transient protein expression has...long-lasting effect, such as Cre- or Cas9-mediated recombination. Both primary hESCs and immortalized cells treated...

...I’ve always been fascinated by the human body and how it works. I used to stay up past my bedtime, poring...baby’s health but adamantly refused to get them vaccinated. A classmate told me that they never finish a...more likely to rate that study as untrustworthy, inaccurate, methodologically flawed, and biased. As a result...information gained from that research remains inaccessible, behind paywalls and in technical jargon that...