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We narrowed to 792 results for: LAT

Showing: 331 - 360 of 792 results

  1. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...can spread between hosts through horizontal (or “lateral”) gene transfer. Horizontal gene transfer can occur...specialized degradative capabilities are often tightly regulated to be active only in the presence of the target...rapid spreading of genetic material through a population of cells. Cryptic plasmids: microbial mysteries...organization doesn’t necessarily reflect genetic relationships or plasmid incompatibility. Plasmids may also...2004). Cloning Vectors Based on Cryptic Plasmids Isolated from Lactic Acid Bacteria:Their Characteristics...

  2. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...following the latest developments in genetics research, their applications and regulatory policies; keeping...traditional and social); and synthesizing and translating all these into social media and blog posts, lesson...symposia, or courses in social, health or policy-related topics, as time permits. Also seek out opportunities...scientific research does not happen in a vacuum that is isolated from the rest of society or devoid of social context...

  3. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...precipitation. All of these methods ultimately isolate the viral particles allowing you to concentrate...be optimized prior to starting. We recommend calculating the ratio of the surface area of the larger vessel...example, while the PEG precipitation method is relatively inexpensive and does not require an ultracentrifuge...titering. Typically researchers will choose a relatively permissive cell line for titering in order to...information provides a great starting point, it may not translate to the target cell line. To overcome this issue...

  4. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...diseases. Additionally, retroviral vectors can regulate gene expression by delivering shRNA or miRNA. ...vector systems Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast...and lentiviral vectors that are used for gene regulation, gene of interest expression, knockout, and screening...Consortium, also known as the Genetic Perturbation Platform, to produce their shRNA library to support functional...genetic power and allows high throughput genetic modulation studies in mammalian cells and tissues. This ...

  5. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...understanding a specific effect related to an experimental manipulation. Common inbred strains include ...in the population, meaning they more closely mimic the genetic diversity of the human population. Outbred...controlled through a drug or small molecule that regulates Cre expression. The most common drug used to induce...

  6. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ... to specific sequences, causing targeted gene regulation. This can be used to activate transcription during...generation” CRISPR activator. While it requires a relatively simple construct, it exhibits modest levels of...activation experiments. In cases of multiplex gene regulation (activating multiple genes at once), however,...construction: it relies on antibody chains, which are relatively large and are not expressed consistently throughout... WA (2013) Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression...

  7. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...crosslinking agent, a signaling ion, and an allosteric modulator. Because of its importance, scientists have developed...understand how this metal ion is trafficked and regulated. In this blog we will explore some of the most...and organs noninvasively and target a defined population of cells (e.g., a neuron subtype) and specific... calcium. It has been applied to mark active populations of neurons in vivo to study circuit activity.... allows scientists to focus on specific cell populations of interest. Many of the calcium sensors described...

  8. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...assembly By Eric Perkins Though it’s easy enough to contemplate DNA in terms of its sequencing, in vivo studies...targeting a specific chromatin feature (e.g. a methylated histone such as H3K27me3), a linker between these... are sensor-actuators, either transcriptional regulators or chromatin-modifying enzymes. The speed and...used to bring cargo, such as protein or DNA, to manipulate function in receiver cells. There are over 40... G-baToN provides versibility in tracking and modulating the interactions between cells.      Tang et...

  9. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    ...OpenWorm project. The main goal of OpenWorm is to simulate C. elegans as a computer model. When I first heard...biology of the worm into a computer model that recapitulates how molecular biology impacts worm locomotion...neuron knockouts so that we can perturb factors related to worm motion. Working on the Cat Microbiome Project... have members looking at how to re-use it as a platform for monitoring homemade microfluidic chips using...using the fantastic optics and fluidics platform it contains. Biotech equipment becomes obsolete amazingly...

  10. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ... After 24 days of phage propagation, Hu et al. isolated xCas9 1.0-1.4 and subjected them to another 72...-targets. Employing xCas9 for transcriptional regulation and base editing Hu et al. also engineered transcriptional... editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages, ...about xCas9. “I’m not 100% sure xCas9 is going to be flat out better than SpCas9. I want everyone to test ...hoping that these answers come sooner rather than later. Once you’ve tried it, let us know how xCas9 is ...

  11. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...often doubles the volume of culture used for inoculation if we get inadequate plasmid DNA yields. Why ...growth conditions, and the material used for inoculation. To further complicate matters, plasmid copy ...may lead to loss of the plasmid, and using older plates could increase plasmid loss or mutation. Let it...these factors are generally, but not perfectly, correlated. Typical growth times for high copy plasmids...smaller cultures, especially those in deep-well plates, require a faster shaking speed for proper aeration...

  12. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...them under a microscope, fractionate samples to isolate particular organelles and their contents, or perform...to the immature color (green or blue) within a population of timer indicates the age of the pool: the higher...of protein concentration. Calibration curves correlating this ratio to the total expression time can therefore... situation where one wants to understand the relationship between the age of a cell, protein, or cellular...nascent tissues and the formation of limbs are correlated with changes in gene expression. In a developing...

  13. Academic vs. Industry Postdocs

    Type
    Blog Post
    Published
    Nov. 13, 2018, 1:22 p.m.
    ...immersed in that environment sooner rather than later can be helpful. Publishing: As a postdoc at AstraZeneca...you might find yourself feeling scientifically isolated if no other postdocs are in your department, and...surrounded by lab members who are all working on related topics, scientific discussion is often part of ...then let them decide what they’d like to work on later. For instance, AstraZeneca posts most of its positions...resources on the Addgene blog Check out all careers related blog posts here Find 9 tips for a successful postdoctoral...

  14. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...bacterial pellet for the remainder of the cultures and later maxiprep only the cultures with intact ITRs. I use...solution. PEG takes several hours on a heated stir plate to dissolve and requires monitoring so that the ...qPCR Protocol for more details. Pro Tips When calculating the titer for your AAV, remember to account for...plasmid, not that I would know about this… A no template control (NTC) should always be included. While...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

  15. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ... vector and your insert, DpnI digest away the templates, and transform the PCR products into E. coli. ...will require a PCR reaction or two to generate templates for the cloning process. Time spent actually doing...restriction enzyme and ligase based cloning is a relatively inexpensive way to get the job done, provided...because the T4 polymerase needed for this method is relatively inexpensive and can be obtained for about $70... or purify DNA fragments from gels. If you’re relatively new to the cloning game, however, or simply looking...

  16. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    Published
    Aug. 29, 2019, 12:50 p.m.
    ...attending the session, and what is their background relative to your research? Tailor your language to your... you may receive. People often ask about the relationship of your work to recently published papers or...audience just that. Explain what you do know about a related topic, and let them know while you appreciate the...collaborate with a group who presented on a topic related to your research? Find a way to get in contact ...affiliation from the poster heading to get in touch at a later time. With these tips in mind, you’ll be geared ...

  17. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...(Lukashev et al., 2016). The protein capsid encapsulating the genome is majorly composed of three proteins...genes. For this you would need to provide a DNA template to repair the double strand break via homology...Cas9/sgRNA and one vector for your specific DNA template. Using this two vectors system, Manuel Gonçalves...This technology therefore represents a potential platform that could be realized via simple switching of...tools can also be used to deliver Cas9 and DNA templates in somatic cells of adult animals in order to ...

  18. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ...I would mainly do over the price of a used car. Later, as I applied for jobs, I knew I should negotiate... the ultimate resolution can involve ideas not related to the actual data collection task but still be...negotiation skills will help you strengthen relationships with your colleagues and supervisors and lead...negotiation skills are key to maintaining and growing relationships with your colleagues within and outside of your... what you want and can also strengthen your relationships with others who know that working with you will...

  19. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ...in mind! This section of our plasmid pages is relatively new, so some of our older plasmids may not have...publication. Addgene makes this easy by providing a template citation for you to use in your Materials and ... Use the Addgene catalog number to easily find template citations for Addgene plasmids you have used in...on the N or C terminus of the transcribed and translated insert. If it is an empty vector with a tag, ...here. Articles Citing this Plasmid: This is a relatively new feature of our plasmid pages, and provides...

  20. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...not narrow enough to distinguish between closely related colors. Furthermore, most FPs  have a broad range...oxygen deprived environment. Recently, a new GFP isolated from the Unagi eel was shown to mature independently...chromophore. This can be from a few minutes after it is translated to a few hours. For example, superfolder GFP ... of how bright is the emission. Brightness is calculated as the product of extinction coefficient and ...College of Medicine. He is interested in everything related to gene expression, particularly at the RNA level...

  21. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...fluorescent proteins, promoters, enhancers, polyadenylation signals, inducible gene-expression sequences...generate the four initial unique constructs and later combine all four constructs into one 15 kb insert...polycistronic containing multiple identical polyadenylation signals or promoters Gibson et al., 2009 ...”) can be readily reused in the assembly of any later construct (i.e.,functional cassettes are recyclable...Neveu PA. MXS-Chaining: A Highly Efficient Cloning Platform for Imaging and Flow Cytometry Approaches in Mammalian...

  22. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    Published
    July 9, 2019, 12:52 p.m.
    ...rather “let’s see how we can make this fun activity relate to STEM.” Figure 1 shows the activities we had ...initial guidelines I received from the teachers, I formulated the presentation as follows (which could be adapted...molecular mechanism, I was careful to keep it relatively simple. I honed the complexity of the talk by...moving along much faster than it can be officially regulated. I did not try to sugarcoat the potential dangers... on a kid’s face when you’ve shown them how to isolate DNA from a strawberry, or demonstrated that they...

  23. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...visualize PPIs that regulate numerous cellular functions, including ubiquitinoylation, kinase signaling,...generate negative control constructs — more on this later. For more on constructing fusion proteins, check...necessary because split FPs can associate, albeit at relatively low frequencies, without being brought into close...visualized makes it amenable to studying PPIs under relatively physiological conditions when compared to FRET...

  24. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ... thing if CRISPR hadn’t made manipulating the genome cheap and relatively easy? Have all of these things...unlikely to be surprised by many of Zimmer’s revelations. The issues he discusses have been making the...before then. Other areas are newer, such as the relatively recent resurgence of “Do-It-Yourself Science”...its own Franklin in Doug Prasher, who originally isolated the GFP gene but was not included amongst the ...

  25. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...future edits. In contrast, since Cpf1 cleaves relatively far away from the PAM, NHEJ might retain the ...confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing Cpf1...throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except when they were present in...exciting both in terms of basic science and translational applications. This discovery of this type V ...we had a lot more to learn about CRISPR biology. Later work, like the adaptation of Cas13 to RNA targeting...

  26. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...usually smallish peptides incorporated into a translated protein. As depicted in the accompanying cartoon... Thus far Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used ...crystallizing it or testing its function. Small ubiquitin-related modifier (SUMO) can help with folding and stabilization...and purification An affinity tag, generally a relatively small sequence of amino acids, is basically a...Combo Consists of a bacterially derived in-vivo biotinylation signaling peptide (Bio), flanked by hexahistidine...

  27. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...non-homologous end joining (NHEJ), HDR occurs at a relatively low frequency, especially in nondividing cells...the cell to use the deaminated DNA strand as a template. To do so, they used a Cas nickase, instead of...repairs the DNA using the U-containing strand as a template, copying the base edit. Figure 2: Cytidine...repair, thus removing these cells from the edited population.  Another way to improve base editing efficiency...improvement in editing efficiency (Koblan et al., 2018). Later directed evolution efforts yielded an editor with...While cytosine base editors often produce a mixed population of edits, ABEs do not display significant A to...

  28. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...peripheral blood mononuclear cells (PBMCs). PBMCs isolated from whole blood present many benefits for the...patients are reprogrammed less efficiently, no such correlation has been found between the age of the patients... amplification in the host cell. This enables relatively high and long-term expression of the reprogramming...using PBMCs as the source of somatic cells. This translates to only about 16 hiPSC colonies per 100 ml of...Blood Cells: An Efficient Protocol Using Serial Plating of Reprogrammed Cells by Centrifugation. Stem Cells...

  29. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    Published
    Oct. 25, 2016, 2:30 p.m.
    ...Secretary of Labor Thomas Perez updated the overtime regulations of the FLSA. The exemption salary was set at ... postdocs are at each of these institutions (populations potentially from 1 to 5761, according to the ...research on these institutions, we can say with relative certainty that just over 40% of current postdocs...for academia Changes are afoot for the postdoc population, and it is unclear how this will all shake out...effects the new laws will have on the postdoc population. In the current funding climate combined with...

  30. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...do a lot of DNA work but never get to actually manipulate the organism they study (unless if you count ...study of diverse organisms. Methods for genetic manipulation still need to be developed for each organism... currently for terrestrial frogs. Cnidarian-Dinoflagellate symbionts (Virginia Weis) Sea anemone Aiptasia...Henry) This black footed slipper snail can be manipulated by injected RNA and can express fluorescent proteins...is developing tools for pan-arthropod genetic manipulation. The ReMOT control system (Receptor-Mediated...
Showing: 331 - 360 of 792 results