We narrowed to 866 results for: FER
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TypeBlog PostPublishedApril 4, 2023, 1 p.m....have risen into yeast expertise! References and Resources References Liu, Wei, et al., From Saccharomyces...applications in food science and biotech due to its fermentation abilities and other useful traits. The “biotech...lists some of the biological commonalities and differences between K. phaffii and other yeast strains. ...aerobe Number of chromosomes 16 3 4 Preferred carbon source Glucose Glucose Glucose, methanol...Applications for K. phaffii There’s a number of different uses for the phaffii-lous strain. Protein expression...peroxisome studies since their peroxisomes will proliferate upon a media change into methanol and peroxisomal...peroxisomal studies on biogenesis, degradation, and proliferation. Genome manipulation K. phaffii’s genome has...
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Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis
TypeBlog PostPublishedAug. 28, 2014, 3:34 p.m....add loading buffer to your digest reactions before loading them. The glycerol in the buffer will make sure...based on the predicted sizes and organization of different features within the plasmid. Restriction analysis...can search NCBI for YGOI to find the particular reference sequence if necessary. The example plasmid on ...fragment having increased intensity to serve as a reference band. The uncut DNA (lane 2) shows 3 possible ...visualize the full size of your construct. Consider buffer and temperature compatibility when digesting with...versions of the endonucleases and/or supply custom buffers as means to avoid this issue. For your gel: ...in the gel well and the dyes provide a visual reference point so you can easily assess how far the gel... -
From Lab Bench to Leadership: Transitioning into Management as a PhD
TypeBlog PostPublishedJan. 21, 2025, 2:15 p.m....skills that were quite different from mine. Learning to bridge those differences was a pivotal moment in...technical problems in the lab, management involves a different skill set: mentoring, setting strategic goals,... day-to-day details. Ignoring individual differencesManaging a team isn’t “one size fits all”. Each team... needs. For instance, I had one colleague who preferred detailed instructions and regular check-ins, which...autonomously. Failing to recognize and adapt to these differences could have led to frustration, micromanagement... particular, served as a great "baby project," offering a supportive environment to build my management...noticed tension between two team members with differing approaches—one detail-focused and the other big-picture... -
Viral Vectors 101: Biosensors
TypeBlog PostPublishedJuly 6, 2023, 1 p.m....imaging for advanced monitoring of different factors, in different or the same cellular location, all ...covered with packaging plasmids! References and Resources References Bhalla N., Jolly P., et al. Introduction...biosensors provide quantifiable readouts for levels of different chemicals, hormones, proteins, etc., that you ...as calcium. This varied component, which we will refer to as the analyte, needs to be specifically recognized...and zinc. These ion sensors can be targeted to different cellular locations or tissues depending on the...acceptor of a FRET pair on actinin will produce different signals based on changes to the structure of actinin...organisms, but AAVs make it possible. In AAV systems, different serotypes can be used to affect or determine viral... -
Antibodies 101: Monoclonal Antibodies
TypeBlog PostPublishedJune 29, 2021, 1:15 p.m....antibodies differ from other types of antibodies? How monoclonal antibodies are made “Monoclonal” refers to ...for your experimental needs. References and resources References Bradbury ARM, Trinklein ND, Thie ...experiments, you may be curious about all of the different kinds of antibodies that are available. One common...heterogenous mixture of antibodies that recognize many different epitopes on the same protein - are more likely...to produce monoclonal antibodies. Genetic drift refers to changes in the nucleic acid sequence of the ...impacts the specificity, avidity, and number of different antibodies produced by the hybridoma. Image from..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P... -
Plasmids 101: Screens vs. Selections
TypeBlog PostPublishedNov. 10, 2022, 2:15 p.m....in. The differences They say the devil is in the details - and that’s where the differences in these... RNAi resources and go fishing! References and resources References Wang, T., Wei, J.J., Sabatini, D....? If you’re thinking, ‘wait, those terms mean different things?!’, you aren’t alone! We’ve noticed a lot...selections The common goals The confusion in differentiating between a screen and a selection isn’t surprising... you would individually treat cells with many different drugs and then perform an assay to look at an ...enhancer/suppressor screens. Selections Selections differ from screens in that only survivable manipulations...experimental question. Now that you understand the difference between the two, it’s time to check out our CRISPR... -
Viral Vectors 101: Optogenetic Tools
TypeBlog PostPublishedJune 27, 2023, 1:15 p.m....excitatory and inhibitory channels activated at different wavelengths of light (Vierock et al., 2021). Step-function...stay on until a second pulse of light, with a different wavelength, turns it off, allowing the cell to... secondary signaling pathways. There are many different opsin (natural and engineered proteins responsive...pathway to activate various protein regulators at different points in the pathway. Photoswitches are great...successive steps along a pathway, there can be very different responses to long vs. short stimulation times ... & Cui, 2015). You’ll need to test for these differences when designing your experiment. Finding the ...available to help light your path! Resources and references Recommended reading Abreu, N., & Levitz, J. (... -
Finding Your Perfect Job After University
TypeBlog PostPublishedJan. 12, 2016, 3:30 p.m....’s of working in different sectors. Luckily, I have worked for a variety of different organizations, so...overview of my experience and how I felt about the different positions to enable them to decide for themselves...effectively. I had to learn how to cope with a different climate and culture, and also needed to be resourceful...very quickly; as soon as I learned how to do the different tests and became efficient, I then started to ...diagnostic tool for the detection and diagnosis of different types of pancreatic cancer and highlighting which...were keen for me to become involved with many different aspects of the company and gave me the freedom... and travel to meet scientists, making my job different every day. I love the freedom to design and develop... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog PostPublishedNov. 6, 2015, 3:30 p.m....international mentoring conference held at the University of Valencia (Spain). At this conference, mentors and mentees...professional development and networking resources offered by programs like IMP USA-Europe not only provide...international mentors. Universities from five different geographical locations in Spain were enrolled:...mentees held scheduled monthly progress video conferences and had real-time correspondence via e-mail throughout...Mentorship Program and the International Mentorship Conference In addition to a pre-program self-assessment ... exchanged impressions of the IMP experience. Conference highlights included remarks by the Spanish Secretary...community overseas. Official representatives from different Spanish universities and high level public and... -
CRISPRainbow and Genome Visualization
TypeBlog PostPublishedFeb. 28, 2017, 3:30 p.m....situ hybridization (FISH) and chromosome capture suffer from low resolution and can’t be used in vivo. ...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three distinct colors...orthologs is key: since each ortholog requires a different PAM sequence, a gRNA designed for one dCas9 should...specific for telomeric sequences and showed that different fluorescently labeled dCas9s are efficiently directed...target sequence. They succeeded in labeling two different pairs of chromosomes using gRNAs specific to sequences...specified by PAM sequence, each color requires a different Cas9 ortholog, and the target sequence must be... green, and red. In contrast, a gRNA with two different hairpins will produce a secondary color cyan, ... -
To Each HIS Own
TypeBlog PostPublishedMarch 2, 2023, 2:15 p.m.... interest is then eluted using a high imidazole buffer that outcompetes the interaction between the protein...contaminating protein (orange). A low imidazole wash buffer disrupts the weak interaction of the contaminating...the protein of interest. A high imidazole elution buffer disrupts the interaction of the protein of interest...step to remove the tag in order to prevent it interfering with protein function. In contrast, the polyhistidine...The concentration of imidazole used in the wash buffer should be high enough to disrupt contaminant-metal... of a protein. Neither the N or C terminus is preferable over the other however, tag placement can affect... it is, and the greater the risk that it will interfere with protein function. Additional Benefits of... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....ganglia in movement disorders. References and Resources References Armbruster BN, Li X, Pausch MH, Herlitze... be the best option. Fluorescent proteins can interfere with fused protein function Under some circumstances...found that the fluorescent proteins can, indeed, interfere with the correct function of the protein of interest...in monkey neurons, we next hypothesized that a different tag fused to the hM4Di might improve the transport...Surprisingly, we found that this small change made a big difference. Replacing the mCherry with the HA tag resulted...transgene of interest. In our studies, mCherry interfered with the transport to the membrane of the DREADDs...doi.org/10.1002/0471140864.ps0909s73 Additional references on the Addgene blog Browse Fluorescent 101 blog... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....there’s less malignancy because there’s less proliferation. However, the lack of caveolin 1 in breast cancer...growth mediators and improving cancer cells proliferation. How do microenvironments affect tumor spread... integrins (cell-adhesion proteins) targeting different organs. This means that exosomes not only promote...microenvironment’s malignancy but also incite different organs to become malignant. Exosomes also contain... (micro RNA) and other molecules that can be transferred to other cells, inducing epigenetic changes in...regulates transcription. Without TKA1, cells proliferate and cannot undergo apoptosis, thus multiplying...issues. Follow her on Twitter @BrbaraPinho6. References Allinen, Minna, et al. "Molecular characterization... -
Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number
TypeBlog PostPublishedSept. 4, 2018, 12:53 p.m....were also tested in different plasmid backbones that varied in size or carried different genes. In these ...expression can make data hard to interpret or can interfere with the precision needed in metabolic engineering...ranging from 3-100 per cell, a stabilized promoter buffered against the effects of copy number on reporter...number. Moreover, gene copy number is affected differently by growth rates when a gene is expressed from...Segall-Shapiro et al. showed nearly no position-dependent differences in gene expression. On the other hand, slow growth...promoters can eliminate variability between the different components. When your pathway needs precise fine-tuning...levels, give these stabilized promoters a try. References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...whenever you hear it (whether at a scientific conference or a karaoke bar), you probably want to sigh....demethylation? Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental...demethylation – tight binding is crucial for dCas9 to interfere with DNMT1 even through replication. The dCas9... CRISPR-based tools for epigenetic editing. References and resources: Additional Resources on the Addgene...Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) ... al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun 9, 597. https://doi.org... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 6, 2021, 12:30 p.m....as they are often quite different from one library to another. They also differ from the CPS process in...examine three things: Aligning the NGS result to a reference sequence to confirm backbone elements. Confirming...deposited plasmids is to align the NGS result to a reference sequence like a known backbone sequence. We’re...through BLAST or by direct alignment to an NCBI reference sequence or a sequence provided by the depositor...synthesized regions that have no publicly available reference sequence or plasmids containing genes with many...features using Snapgene. If we find information that differs from what we would expect given the data provided...the depositing laboratory confirms that these differences are expected and do not affect plasmid function... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog PostPublishedDec. 1, 2015, 3:30 p.m....of the Canton Nucleic Acids Forum (CNAF) 2015 conference invited me to attend and speak about Addgene’...Thoughts on Guangzhou and the Science City The conference was located in the modern, relatively new “Science...national science and technology park, it enjoys preferential governmental policies for a high-tech development... which I could identify. There were dozens of different mushroom varieties alone. At least I could tell...requires an interdisciplinary team. Day two of the conference was kicked off by Nobel Laureate (and Addgene...circRNAs do yet or how they do it. Of course, No conference would be complete without CRISPR Michael Terns... also exploring the use of cas9 proteins from different species for genome engineering. The final keynote... -
Important Considerations When Using AAVs
TypeBlog PostPublishedJune 13, 2017, 2:30 p.m.... This preferential tropism for different brain regions is largely due the ability of different serotypes...tissue? What is commonly known is that different serotypes have different tropisms but, as you can see, AAVs...there are marked differences among serotypes in their efficiency at transducing different brain regions....serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction...movement. Follow her on twitter @katrinaboards. References 1. Russell, David W. "AAV vectors, insertional...therapy 8.16 (2001): 1248. PubMed PMID: 11509958. 8. Ferrari, Forrest K., et al. "Second-strand synthesis is... -
9 tips for a successful postdoctoral experience
TypeBlog PostPublishedOct. 9, 2018, 12:21 p.m.... NIH, in the United States) Strong letters of reference that attest to your talent and expertise as a ...Leader position applications require 3 letters of reference- the graduate school mentor, the postdoctoral ... and your research well. Who could fill these different roles? A second or third mentor. Therefore, seek...the job market. Additional mentors are not so different from having a thesis committee in graduate school...confidence in the reagents and outcomes. This is preferable to finding out you've been doing something all...Finally, if you give your talk or poster to enough different kinds of audiences and if you pay attention to...presentations and will be able to provide letters of reference, informal recommendations, more positive grant... -
Antibodies 101: Validation
TypeBlog PostPublishedMarch 24, 2022, 1:15 p.m....discussions on strategies for antibody validation in different applications. Here, I want to highlight the general...transcriptomic or proteomic data from several different tissues or cell types. If your antibody indeed...antibodies against the same protein, each targeting a different epitope, and look for correlation between the ...the results. Using antibodies against different epitopes on the same target reduces the likelihood that ...unique antibodies. An additional caveat is that different antibodies may or may not perform well under the...couple of antibodies that are predicted to bind different epitopes of this protein. You perform flow ...similar results, while the third looks entirely different (B), giving you confidence that your new antibodies... -
PCR Applications for Copy Number Variation Assays
TypeBlog PostPublishedApril 26, 2022, 1:15 p.m....several different markers, it can also be used to run multiplex assays, allowing the reference sequence...primer Molecular Biology Reference Molecular Cloning Techniques References Bell AD, Usher CL, McCarroll...each cycle. The raw data is then normalized to a reference sequence present in the same amount in all samples...curve. For CNV assays, it is recommended to use a reference sample with two copies of the gene (Ma, 2014.)...probe set for both the gene of interest and the reference sequence. At the end of the run, each droplet ...positive droplets for the gene of interest and reference gene, copy number can be determined through concentration...that matches your needs. Happy quantifying! References and Resources More Resources on the Addgene blog... -
Genome-wide Screening Using CRISPR
TypeBlog PostPublishedAug. 18, 2015, 2:30 p.m.... are composed of many different gRNA-containing plasmids, each with a different genomic target. The goal...expressing cells. Pooled lentiviral CRISPR libraries (referred to simply as “CRISPR libraries”) consist of a ...heterogeneous population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within.... Pooled gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries...cell or set of cells containing a mutation in a different gene. Libraries may be available in a 1-plasmid... be used in drug screens to identify genes that confer drug-resistance. Mutant cells are treated with ...gRNAs that are “enriched” correspond to genes that confer drug resistance when mutated. Findings from this... -
Why Do I Need an MTA Anyway?
TypeBlog PostPublishedMay 1, 2014, 8:23 p.m....Uniform Biological Material Transfer Agreement (UBMTA) for its material transfers. The UBMTA was drafted by... has time for paperwork? Here’s why a Material Transfer Agreement (MTA) matters: Science thrives on collaboration...universities and institutions require them for material transfers. When Addgene entered the plasmid sharing space...or choose to auto-approve MTAs. Whatever your preference, we are here to make it fast and easy. Fast...receiving institution. Therefore, many technology transfer officers are familiar with the UBMTA and are happy....org or +1 617-225-9000, or check out our MTA reference pages. So go ahead and use Addgene plasmids ... material transfer agreement... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog PostPublishedDec. 2, 2014, 7:23 p.m....about your different experiences in research labs. What are some similarities and differences you have ...found that research institutes in different places have very different cultures. In Basel, I felt that ...recently developed the oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) method to suppress PCR amplification...join the Advisory Board of Addgene, I accepted the offer without any hesitation. I hope I will be able to...world? Fujii: Fortunately, I was able to work in different parts of the world in my early research career...technologies, hopefully they'll be completely different from what we are now doing in my lab then when...to Alerts" link in the Addgene Alerts box. References Identification of Proteins Associated with an... -
Which Fluorescence Microscopy Technique is Best for Me?
TypeBlog PostPublishedOct. 10, 2017, 1:57 p.m....opposite direction to fluorescence collection is referred to as “Epi” illumination. Therefore these are ...incorporate optical sectioning. Optical sectioning refers to the ability to extract light from a single, ...excitation light path. Each hole corresponds to a different location in the sample. As more than one hole ...between the glass coverslip and a sample in aqueous buffer it is completely reflected. This reflection occurs...index mismatch between the glass and the water-like buffer the sample is immersed in. Although the excitation...one of four categories that are each suited to different types of modern day fluorescence microscopes (...means that the images will appear sharp without interference from blurry out-of-focus light. The most difficult... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog PostPublishedNov. 21, 2024, 10:47 p.m....many countries, import and export regulations, differing customs processes, and the distances the packages...distributes to per year, 2005-2023. We identified and offered a number of solutions that increased accessibility...each courier we contracted with so we can always offer the fastest shipping. However, there were issues...specific to that country. For example, when we began offering temperature-sensitive materials, we noticed a ... backbones, lentiviral packaging systems, and luciferase expression. On individual material pages, we ...educational resources for each type of resource we offer. In 2013, we launched the Addgene blog for topics...particular topic into beautifully designed and easy-to-reference eBooks. Figure 3: Global readership of the Addgene... -
Hot Plasmids - November 2023
TypeBlog PostPublishedNov. 14, 2023, 3:35 a.m....optimized for dual-AAV systems. These variants use different RT enzymes and performed large edits with similar...types. DIRECTED is compatible with fusogens from different viral families and can be used to target multiple... to reduce negative side effects. This platform offers a promising approach for experimental therapies... enzymes, TurboID and APEX2, are localized to different cellular compartments. They showed APEX2 can use...uses sequential labeling by TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled...stably or inducibly express tagged iPAK4 from different promoters, specific time points appear as fluorescent...bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record transcriptional... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog PostPublishedNov. 10, 2016, 3:30 p.m....acquired drug resistance are referred to as methods of ‘horizontal gene transfer’. Resistance can also arise...you! Many thanks to our Guest Blogger, Jennifer Rivers! Jennifer Rivers ScM, MLS(ASCP)CM is a research ...This post was contributed by guest blogger, Jennifer Rivers, a research microbiologist specializing in...multi-drug resistance are resistant to three or more different classes, or types, of drugs used to treat them...either through random mutation or horizontal gene transfer, means to defend themselves from these naturally...resistance can arise, and has arisen, for many different reasons. Exposure to antibacterial drugs provides...science communication, and public health. References 1. O'Neill, J. "Tackling drug-resistant infections... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog PostPublishedJuly 23, 2019, 12:46 p.m....strains of mice. This suggests that a small genetic difference between these two mice might drive their divergent...took a broad approach and searched for genetic difference between the 36 mouse strains that are part of...permissive phenotype, but only one, Ly6a, showed differential protein and RNA expression between the brains...are not reproduced when the vector is used in different mouse strains. The in vivo selection for PHP.B....eB viral preps are available from Addgene. References Holmes, Christina, and William L. Stanford. "...performed in non-human primates, check out these two references: Dayton, Robert D., Mychal S. Grames, and Ronald...Ronald L. Klein. "More expansive gene transfer to the rat CNS: AAV PHP. EB vector dose–response and comparison... -
Supporting Rare Disease Research with BeHeard 2017
TypeBlog PostPublishedJuly 20, 2017, 2:30 p.m....patients suffer any less acutely. The Rare Genomics Institute works to help those who suffer from rare...in diabetes and lymphoma research or the many different ways you can improve your heart health. These ...disease research and provide hope to patients suffering from these devastating afflictions. Elia Di Schiavi...abdomen and affects roughly 20% of mesothelioma sufferers. This disease is less widely understood and often...for peritoneal mesothelioma and thereby relieve suffering from this little-known disease. You can find more...Nabavi and will continue to work to help those suffering from rare diseases. These underrepresented individuals...rare disease research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog PostPublishedMarch 5, 2019, 1:08 p.m....through the proliferation of specific, and not all, Langerhans cells, to create foci of proliferating cells....construct containing fluorescent proteins (XFP) of different colors to label an organ or entire organism. Because...more refined versions have been developed, which differ in the number, stability, and brightness of the...localization sequence, allowing it to localize to a different area of the cell than CFP and YFP. The most recent...could recombine to activate the expression of a different fluorescent protein. Therefore, each cell could...become a powerful tool for understanding cell proliferation. Inducible XFP expression in the Ubow mouse ...can be combined with new imaging techniques. References Cai, Dawen, et al. "Improved tools for the Brainbow... -
Evolution of Lab Techniques
TypeBlog PostPublishedJune 21, 2016, 2:30 p.m....through a tiny pore by measuring differences in ion flow caused by the different electrochemical properties ...fragmented into smaller pieces, run on a gel, and transferred to a membrane for analysis through hybridization...standard PCR. This allows the researcher to compare different DNA sequences to see which is more abundant in...content by comparing the fragments created by different restriction enzyme digests. Incubating a DNA sample...restriction mapping (Figure 1). Though some labs may prefer to send DNA directly for sequencing (more on that...incorporation of a ddNTP. Researchers perform four different reactions, one for each radiolabelled ddNTP, and...machines (9). Today, similar technologies, sometimes referred to as sequence DNA as it synthesized by electronically... -
Plasmids 101: A Brief History of Plasmids and an Improved eBook!
TypeBlog PostPublishedOct. 29, 2015, 2:30 p.m....understand genetic cytoplasmic factors that could be transferred between cells. At the time, these extranuclear...episome seemed appropriate, especially since the Fertility, or F-factor discovered by Ester Lederberg in ...-factors. Like F-factors, R-factors could be transferred between bacteria via cell-to-cell contact; however... the same. Over the next few years genes from different bacterial (and eventually mammalian) species were...ubiquitous tool that can be diversely applied to many different experiments. Addgene was founded in order to store... keeping with our goal of creating a one-stop reference guide for plasmids we've updated and added new...: 24043817. PubMed Central PMCID: PMC3785787. References: Lederberg, Joshua. "Cell Genetics and Hereditary... -
Interview: Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering
TypeBlog PostPublishedMarch 4, 2014, 4:49 p.m....specific problem. Every species is so different – an awake monkey is different from a cell in a dish. I always...of life. Our most recent paper in Nature Methods offers the first true two-color pair. We'll be sending...want to activate two sets of neurons and see how different parts of the brain or the cells within them work...set of neurons, maybe we could treat patients suffering from Parkinson's, chronic pain, deafness, or other...Boyden: I think because neuroscience has so many different goals: to understand how a mouse changes emotions...find ways to make these tools more selective for different ions. Right now, we can modulate electricity. ... click the link in the Addgene Alerts box. References: Independent Optical Excitation of Distinct Neural... -
Addgene Depositors Get More Citations
TypeBlog PostPublishedApril 20, 2017, 2:30 p.m....deposits to Addgene. Insomuch as different journals represent different research areas, target readers,... would also be great to know how much of that difference in citations is caused by depositing a plasmid...wouldn’t be causal. To control for a number of differences between those papers with Addgene plasmids and..., so our estimates now take into account any differences in citations due to the journal of publication... the same journals! So, is all this remaining difference caused by depositing the related plasmids with...it is still suggestive that we see such a big difference in citations after controlling for journal. And...Visit his website at: http://www.neil-t.com/ . References 1. Cong, Le, et al. "Multiplex genome engineering... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog PostPublishedApril 18, 2019, 12:53 p.m....cells (Ma et al., 2015). To do so, they fused a different fluorescent protein to various Cas9 variants, ...either MS2, PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to the ...green, or red readout. But gRNAs containing two different hairpins will produce a secondary color (yellow...allows for simultaneous visualization of sites on different chromosomes in living cells, the system requires...they chose an intergenic DNA region (IDR3) as a reference loci to measure inter-locus distances of loci ...sgRNA-Sirius-4X(MS2-PP7) showing recruitment of different fluorescent proteins. (B) CRISPR-Sirius used to...try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR labeling... -
What Good Citizenship Can Do for Reproducibility in Science
TypeBlog PostPublishedNov. 7, 2019, 2:18 p.m....research program (think a subtle difference in mouse chow, a different water filtration system, environmental...outline and refer back to the original paper that everyone in your lab has been referencing for the last...original researchers actually did. Admittedly, the differences are often very subtle, and it can take a lot ...and it was just that they were doing something differently. Communication comes in many forms—from transparency...other publishers and organizations have various different versions of checklists and guidelines to help ...rewarding good scientific citizenship in all its different forms as part of graduation, hiring, funding, ... -
R Bodies: Membrane-Rupturing Microscopic Tools
TypeBlog PostPublishedApril 14, 2016, 2:30 p.m....not exert force by dynamically rearranging into different polymerization states. Instead, R body monomers...natural context or 3) be modified to extend at different pH levels. Domesticating R bodies We have shown...that R bodies can function to break membranes of different organisms. And finally, we demonstrated that R... can be “tuned” with mutagenesis to extend at different pH values. Using a screen that takes advantage... fact that coiled R-bodies sediment in certain buffers but remain soluble when extended (see figure to...region produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild ...School. Follow her on Twitter @jessicapolka. References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond... -
Components of CRISPR/Cas9
TypeBlog PostPublishedFeb. 2, 2016, 5 p.m.... labs (see table below) have different PAM requirements. These different PAM requirements allow researchers...discovered all the time and the requirements of these different systems can vary (for example, xCas9 is a variant...neither bind nor cut the target. Cas9 homologs from different organisms or Cas9 mutants developed in a variety...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...). Throughout the remainder of this post, we’ll refer to this RNA complex as the “gRNA”. The gRNA contains...choice (see tips for designing a gRNA) and their prefered Cas9 variant (with the appropriate PAM sequence...sequence. Now that you know about all the different components of CRISPR, It's time to start thinking... -
Advice for Scientists Starting a Lab
TypeBlog PostPublishedApril 29, 2014, 1:43 p.m....student or postdoc may require different types of mentoring. Some mentees prefer lots of advice and micromanagement...an effective working environment? Tom Ellis was offered a start-up package that included a postdoc and ...recommends to seek out advice from seasoned PIs who can offer support. “It is an exciting and emotional time!”...recruit people who are nice as top criterion and offers advice to not rush when growing your lab. “One ... lab and safety manager, researcher, adviser, conference planner, and writer.” Connie Cepko was surprised...impossible' or useless.” How do you run your lab differently or the same as your adviser did? Tom Ellis merges...online. The Howard Hughes Medical Institute (HHMI) offers some great resources for Early Career Scientists...