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We narrowed to 842 results for: Eras

Showing: 321 - 340 of 842 results
  1. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...carbon sources such as glycerol or glucose (in moderation) may also serve to increase cell density and ...plasmid yield. You can optimize your growth time, temperature, and shaking speed to maximize cell density and... and plasmid yield as these factors are generally, but not perfectly, correlated. Typical growth times...sufficient gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around...plates, require a faster shaking speed for proper aeration (260-300 RPM). 37℃ doesn’t work for every plasmid...plasmid Certain plasmids and strains grow best at temperatures other than the standard 37℃. This plasmid feature...recommendations to grow a plasmid at 30℃ or room temperature. These recommendations also often require a longer...
  2. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    Published
    Dec. 12, 2013, 3:35 p.m.
    ...describe and interact with the world using words like OR and AND. These logical operators are so useful...collection is the beginnings of what we think of as an operating system for engineering an entire cell. Typically...BIOFAB project is seeding the beginnings of an operating system for programming life, starting with the... pieces in these kits as the beginnings of an operating system or language for programming life. Which...as applied to products on the domestic market generates $300 billion dollars every year. That's two percent...anticipate plugging into the probiotic and cellular therapeutic space. If you were to look forward to the future...
  3. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...be stabilized by monomeric superfolded GFP and averages only ~50% RNA knockdown. In initial tests, Cas13b...Prevotella sep. P5-125 (PspCas13b) yielded 62.9% average knockdown, and they chose this enzyme for further...editing. To increase A->G editing, they employed hyperactive ADAR constructs, like ADAR2DD(E488Q). They also...and designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found...off-target activity. By mutating residues in ADAR2 that interact with duplex RNA, they hoped to destabilize ADAR-RNA...   Addressing the potential use of RESCUE in therapeutic applications, the authors tested various dRanCas13b...of RNA editing in research and its potential therapeutic uses We are also curious if REPAIR and RESCUE...
  4. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    Published
    July 16, 2020, 1:15 p.m.
    ... to illuminate biology and enable next-generation therapeutics. Prime editing, base editing, PACE, and... lab places an order, they are likely to order several plasmids at once, breaking the statistical independence...our distribution, resulting in the following hierarchical model: Because the U.S., Europe, and China ...the inferred percentage of normal activity (the average rate of unique PIs/labs placing orders per day)...The inferred weekend effect is, in retrospect, generally more dramatic than the COVID-19 effect, though...not a causal interpretation -- the effect would generally not be the same if weekends were somehow imposed...
  5. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...fresh columns, regenerated columns had similar DNA yields. Additionally, columns regenerated up to five times...size of bacterial culture you’re working with. Overall, the glass syringe filter method is not only cheaper...cellulose dipstick. Image: Zou et al., 2018. Regeneration of silica columns for DNA purification Starting... and Deng labs developed a simple method for regenerating silica columns that come with PCR product purification...before a wash with ddH2O and then ethanol. After regeneration, trace amounts of DNA (3 femtograms/ul) could...Huang, L., Zhao, S., & Deng, W. (2018). Rapid Regeneration and Reuse of Silica Columns from PCR Purification...
  6. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...your viral vector, you use the same protocol for generating lentivirus, but you use a different envelope ...terminals and can’t be used for neuronal tracing or peripheral gene delivery. Combining the rabies glycoprotein...Cells Are Inactivated by Human Serum. Molecular Therapy 2:218–222 . https://doi.org/10.1006/mthe.2000.0116...Modification of Hematopoietic Cells for Research and Gene Therapy. Viruses 12:1016 . https://doi.org/10.3390/v12091016...Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods 28:291–301 . https://doi.org/10.1089/hgtb...transport and access to the nervous system after peripheral delivery. Human Molecular Genetics 10:2109–2121...envelope glycoprotein show increased stability in sera and augmented transduction of primary lymphocytes...
  7. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...base-pair mCherry insertion. Editing of these motifs generates lesions that are subject to excision and ssDNA...This edit changes the histidine to a tyrosine generating a GFP variant (GFPY66) that has a shifted emission...Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image...hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that contains an ACG start... of different reporters including mScarlet-I, luciferase, antibiotic resistance markers, and other enzymes...fluorescent proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor AC, Rees HA,...CRISPR/Cas9-mediated Genome Editing. Molecular Therapy - Nucleic Acids 5:e334 . https://doi.org/10.1038...
  8. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ... the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the General Meeting ...the next generation of scientists. The connection between mentors and Singletons can accelerate the networking...meet someone new, as they’d been introduced to several people by their mentor, as well as their bin-mates...recognized by meeting organizers in their town hall.  Iterative improvement - Making Binning Singletons better...Goldschmidt conference, Coastal & Estuarine Research Federation (CERF) Biennial Conference, Gordon Research Conference...
  9. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    Published
    Jan. 23, 2020, 2:45 p.m.
    ...process to reduce viral order fulfilment from an average of 40 minutes to 10 minutes. While users can’t ...result in less than optimal user experience and the overall soundness of the software design. Each of us brings...called Scrum. The Scrum framework is based on an iterative and collaborative approach to development. Scrum...capacity to finish all of the work. Since we work in iterative chunks, we have the flexibility to adapt and easily...allows for life to happen without impacting the overall work. Each of our sprints is themed and the stories... being maintained (i.e. hasn’t been updated in several years) or they make significant changes that make...that helps our users focus on the fun stuff, accelerating research and discovery. We work with transparency...
  10. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, that overcome...delivery and reduce off-target effects of gene therapy, Feng Zhang’s lab has developed a system called...specific cell types. It can also make CAR-T cell therapy more accessible by simplifying the production process...creation of CAR-T cells directly within the body. Overall, DIRECTED can increase on-target delivery with ... offers a promising approach for experimental therapies and medical applications.  Find DIRECTED plasmids...Lacy Proximity labeling can identify protein interactions or map a local proteome, but some applications...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into...
  11. Lab to Office Culture Shock

    Type
    Blog Post
    Published
    Feb. 5, 2019, 1:04 p.m.
    ...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills... 5 o’clock. You’re going to have to talk and interact with...non-scientists! Maybe you’ve just spent... Wu in graphic design. You could dread these interactions, but you could also embrace them. Talking about...production, I didn’t really start to recognize my transferable (or “soft”) skills until I was out of the lab...lab to think about these--get started with our Transferable Skills Guide.     Over the last decade, my... blog Visit our careers topic page Read our transferable skills guide Read blog posts about career readiness...
  12. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ... slim chance of generating indels with base editors, keep in mind that the generation of indel sometimes...minimizes the second outcome and increases the generation of the desired T-A base pair. Figure 1...inosine), which is treated as ‘G’ by cellular polymerases. Subsequently, an original genomic A-T base pair...simultaneously.     These binary vectors could be used to generate stable genome edited plants via Agrobacterium-...transformation. Cytosine base editors for plant There are several CBEs available for plant systems. Target-AID based... transient assay. For stable transformation, regenerated plants may be subjected to genotyping. If successful...
  13. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...India at room temperature. Partnering with our distributor allowed us to ensure that temperature-sensitive...transit for a certain number of days and tracking average transit time for each courier we contracted with...destination in a timely manner and at the correct temperature. To date, we have partnered with six distributors...country. ‌For example, when we began offering temperature-sensitive materials, we noticed a large percentage...preps and libraries were kept at the appropriate temperature for the duration of their journey. Figure 2... backbones, lentiviral packaging systems, and luciferase expression. On individual material pages, we ...
  14. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ...and some considerations when using the method. ELISA basics All ELISAs follow the same overall process....simple and saves time, it also has several drawbacks. Direct ELISAs generally have higher levels of background... the capture reagent to prevent non-specific interactions. Next comes the binding step where the target...Typically, the reporter is an enzyme such as horseradish peroxidase (HRP) or alkaline phosphatase (AP)... of conjugated antibodies   Additional considerations No matter what ELISA method you choose, it ...
  15. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...initiation proteins to start their own replication. In general, these plasmids tend to be low copy number. Conversely...at the low-affinity sites, the proteins work cooperatively to separate nearby AT-rich regions of DNA. In...are occupied by ATP-bound DnaA, the proteins cooperatively can separate the DNA strands. Created with BioRender.com... from the ori. In the elongation phase, a DNA polymerase synthesizes a new DNA strand at each replication...controlling the initiation of replication. Once the average copy number for a given plasmid has been reached...called iterons located near the ori and directly interacts with DnaA, stabilizing its binding and contributing...negatively regulate its own expression and thereby the overall replication of pSC101 (del Solar et al., 1998)....
  16. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...means they can be used with or as fluorophores in several different ways. Researchers have used oligonucleotides...can be created in many different ways, but the general premise is that an aptamer in its unbound state...cells. The new aptamer also has a higher melting temperature than its Spinach ancestors, and has a lower magnesium...fluorophores, including: Environments with higher temperatures, since proteins can denature while aptamers ...likely to trigger immunoresponses. Nucleotides, in general, are far less immunogenic than proteins. Assays...ions or particularly small molecules that do not generate an immune response. Aptamers can be selected for...Peixuan AND Haque, Farzin. RNA Nanotechnology and Therapeutics. Boca Raton: CRC Press, 2014. 5. Hayat, Akhtar...
  17. Interview with Bart Newland, Addgene’s Newest Board Member

    Type
    Blog Post
    Published
    Aug. 3, 2021, 1:15 p.m.
    ...company that is developing RNAi therapeutics to treat neurodegenerative diseases. He has earned a B.S. ...s Board of Directors, Bart Newland. Newland is General Counsel at Atalanta, an early-stage biotech company...like? BN: I've made the transition now to being general counsel of a small start-up. For the last 20 years...and Biogen. They’re very different jobs. Being general counsel at a small start-up is very interesting...interesting. It’s very varied. There’s IP work to do, general corporate work, and negotiating contracts with other...
  18. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...of organelles and vesicles, and more. Although several strategies are available to disrupt microtubules...extended to other GTPases. The Chow Lab has also generated a variety of membrane-recruitment optogenetic ...recently developed a full toolbox and workflow for generating new Rho GTPase optogenetic systems (Berlew et...express the other separately, then use blue light to generate CRY2-CIB1 oligomers that trap the fusion protein..., and 100 times the activity of the original, generated using directed evolution (Tei et al., 2023).  ...license.   Just make sure you don’t overdo it! The moderately-enhanced 10x and 30x SuperPLDs avoid dark-state...Krishnan, N., Ononiwu, F., Pal, D., Manikas, J., Taveras, C., Hall, N. A., Silva, J. D., Freshour, J., &...
  19. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ... genes with potentially harmful mutations in therapeutic settings. Silencing of a target gene is achieved...RNA (shRNA) Short interfering RNA (siRNA) Piwi-interacting RNA (piRNA) miRNA and piRNA can be found endogenously...experiments. shRNA plasmids typically use RNA polymerase III promoters, which transcribe small RNAs. The..., shRNA plasmids are being designed using RNA polymerase II promoters. The design of these plasmids is...plasmids One of the most common uses of shRNAs is to generate stable knockout or knockdown cell lines. In fact...collections/mammalianrnai/","type":"EXTERNAL"},"user_generated_content":false}{% endraw %}{% end_module_attribute...(2004). MicroRNA genes are transcribed by RNA polymerase II. The EMBO Journal, 23(20), 4051–4060. https...
  20. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...molecule into the cell. The fluorescence that is generated allows for qualitative and quantitative measurement...contact with its RNA aptamer. This increased interaction between Corn and its small molecule makes Corn...developed to quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts... transcripts uniquely transcribed by RNA Polymerase III (5S, tRNA and U6) and measuring Corn fluorescence...effect of mTOR inhibition on the activity of RNA Polymerase III. Red and orange fluorescent aptamers Corn...changes in intracellular viral rna." Journal of General Virology 98.5 (2017): 992-1003. PubMed PMID: 28555544...PMC5602116. Song, Wenjiao, et al. "Imaging RNA polymerase III transcription using a photostable RNA–fluorophore...
Showing: 321 - 340 of 842 results