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We narrowed to 811 results for: LARS

Showing: 241 - 260 of 811 results

  1. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ...Now, scientists are beginning to see Addgene as a large data set. Addgene has over 65,000 plasmids in the... in synthetic biology. “Synthetic biology, in particular, the ability to do DNA synthesis, is evolving...protein, their gene sequence may have slight or large variations depending on how closely related these...within the Addgene repository is mammalian, but the largest source of unique gene sequences is Proteobacteria...the only time researchers have used Addgene as a large data set. Alec A. K. Nielsen & Christopher A. Voigt... page Search for a plasmid Find Addgene's most popular plasmids ...

  2. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ...Repeating sequences within a plasmid can undergo intramolecular DNA recombination. This naturally occurring...recommendations from the depositing lab. If your vector is large (>10 kb) or has a low copy number, try following...agar plates, you may notice that there are both large and small colonies. Bacteria containing the recombined...plasmid, whereas the round tan colonies that look similar to normal E. coli cultures contain the recombined...: electron microscopic analysis of in vitro intramolecular recombination. Proceedings of the National ...Additional resources on the Addgene blog Find more molecular biology and viral vector protocols Learn about...

  3. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    Published
    Oct. 10, 2019, 12:41 p.m.
    ...Stability improvement of antibodies for extracellular and intracellular applications: CDR grafting to stable...image nascent proteins. In addition GFP tags are large, dim, and permanently attached to a protein, making... detects the widely used HA epitope in diverse cellular environments. They called this probe the HA frankenbody...else can you do with the HA frankenbody? Another large advantage of the HA frankenbody is that it remains... fluorescent protein guide Find plasmids for subcellular localization Find fluorescent protein plasmids...

  4. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    Published
    Sept. 26, 2017, 12:12 p.m.
    ...unruly. Instead of settling down into a singular focus on molecular pathways, I found myself curious about... a look at the blog ‘Scholarly Kitchen’ for a glimpse into the world of scholarly publishing. And see ...programs unfold, absorbing the stochasticity of molecular interactions and embedding individual diversity...viewed through the lens of development, cell, and molecular biology. Beyond that, she is also a fan of baseball...

  5. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    Published
    April 7, 2015, 4:21 p.m.
    ...eukaryotic cell for the execution of many complex cellular behaviors, such as differentiation or metabolism...metabolism. Our attempts to manipulate these cellular behaviors often fall short with the generation of various...control, a new study repurposes the sgRNA to include modular synthetic RNA scaffolds that can recruit various...master ON/OFF switch. This will likely prove particularly useful in avoiding toxic intermediate buildup...technique that it should be on the radar of every molecular microbiologist. Summary  The single guide RNA...the CRISPRi system can be repurposed to include modular RNA scaffolds. These RNA scaffolds can recruit...

  6. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ..., which can be done by isolating animals with particular visible phenotypes. These approaches may become...) with 30-60 basepair homology arms to knock in large epitopes, such as GFP (Paix et al., 2014). A further...my initial direct screening efforts, which were similar to the approach detailed by Paix et al. Although...Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of a zen-4(cle10ts...facilitated CRISPR editing in C. elegans. These modular toolkits build single plasmid targeting vectors...the “elegans” methods outlined in this post – particularly the use of co-selection methods – could greatly...Tzur YB, Friedland AE, Nadarajan S, Church GM, Calarco JA, Colaiácovo MP (2013) Heritable Custom Genomic...

  7. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...academia, industry, and especially the clinic. Popular validation assays are insufficient As discussed...; Pattanayak et al. 2013). This approach misses larger mismatches (e.g. six nucleotides) that may still...require high read counts (fast multiplexing) Requires large cell population, senstive to time since cell fixing...We additionally thank Monica Sentmanat, Victor Dillard, and Ayokunmi Ajetunmobi for their contributions...diagnostic and clinical implications." The Journal of Molecular Diagnostics 12.4 (2010): 425-432. PubMed PMID:... TALENs, CRISPR/Cas9, and megaTAL nucleases." Molecular Therapy (2015). PubMed PMID: 26502778. 13. Frock...Optimizing CRISPR-Cas9 Genome Editing Specificity." Molecular Cell 63.3 (2016): 355-370. PubMed PMID: 27494557...

  8. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ... TF ORF. From this large sequencing endeavor, a map of possible TF-induced cellular states was generated...both up- and down-regulated pathways. Individual cellular fates were then matched with candidate TFs responsible...an invaluable tool to be used in this capacity. Similar to the Human TFome Library, the MORF library can...allowing for combinatorial overexpression programs on cellular processes. For help with library transformation...expression control systems to mimic illness on a molecular level. Atlas resource The TF map is a valuable...

  9. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    Published
    July 25, 2023, 1 p.m.
    ...Addgene.org Addgene's most popular lentiviral plasmids Addgene's most popular retroviral plasmids Addgene's...organisms. This is often cDNA, shRNA, or something similar, and is contained on the transfer plasmid. If expression...enveloped which is achieved by expression of the vesicular somatitis virus G glycoprotein (VSVG). VSVG is...different from gamma-retrovirus, but they perform very similar functions to infiltrate the host cell and deposit.... Viral packaging and entry Adenovirus is quite large and has more helper plasmid components than the ...

  10. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...have been derived from. Lentiviruses have slightly larger packaging capacities than gamma-retrovirus, coming...closely related, their required components are very similar. While plasmids containing virus-specific elements...currently known to cause any diseases. Recombinant AAV largely does not integrate, and the addition of a helper...to specific cell types of interest. This is a particularly nice feature as organ-specific injections, or... that of lenti and gamma-retro viruses. This is large enough for many applications, but in some cases ...transient expression of a genetic cargo or have a very large cargo. Fast fact: All viruses discussed in this...

  11. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced... available from Addgene. Another way to dodge cellular variation observed in mammalian cells is to tag... all experimental conditions into account. In particular, the folding and maturation efficiency of the...Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced...Marieke Mastop is a PhD candidate at the Section of Molecular Cytology (University of Amsterdam) where she develops...Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein...

  12. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ...techniques have become indispensable tools for molecular and cell biologists over the last two decades,...light. Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be...fused fluorescent protein. This technique is very similar to the widely used FRET, but it eliminates the ...phototoxicity, they successfully imaged multiple subcellular compartments over several minutes using Nano-...concentrations of the compound can be added to the media. Cellular esterase then converts diacetyl coelenterazine-h...important to verify that this substrate does not alter cellular physiology in your system. Optical sectioning...

  13. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs, ...mammalian system. Since Cas13d is functionally similar to previously discovered Cas13 enzymes - what makes...to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But these...their cleavage pattern: EsCas13d produced very similar cleavage products even when guides were tiled across...from the genome, and showed success in models of muscular dystrophy. In this case, Konermann et al. targeted...diversity of CRISPR systems, only time will tell how large the genome and transcriptome engineering toolbox...

  14. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...therefore determines the timing of gene expression and largely defines the amount of recombinant protein that ...vectors. Negative inducible promoter pBad is another popular prokaryotic promoter often used for bacterial protein...induced promoters; they show near-zero expression at regular temperatures but can be induced by heat or cold... transcription to take place. Addgene also has popular light-regulated two component systems designed ...between cell types or organisms, promoters are similarly variable. Bacterial promoters only work in prokaryotic...related species from which they were derived. Similarly, the various eukaryotic cell types (mammalian,...

  15. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...for plasmid cloning Find our molecular biology reference here Find molecular biology protocols here ... or eukaryotic cells is a common technique in molecular biology and scientific research. There are multiple...material is incorporated into the host’s genome at a particular integration site. The integrated phage remains...material can pass through. If you are starting any molecular biology experiment check out Addgene’s protocol... protocols and videos for techniques in basic molecular biology, plasmid cloning, and viruses. References...

  16. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...for copy and paste functionality. Sequence Similar to the plasmid maps, the linear sequences hosted...hosted here are now interactive and have similar hover over features. As with the map tab, clicking on two...enzymes by clicking on column heads. Clicking on a particular restriction enzyme here also highlights it in...standard GenBank feature types. Clicking on a particular feature here also highlights that feature in ...locations, lengths, and direction. Clicking on a particular primer here also highlights it in the “Map” and...Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN...

  17. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    Published
    Feb. 18, 2014, 4:22 p.m.
    ...ends, some 50% of these groups continue to meet regularly, providing ongoing support, advice and accountability...and mentees. Because the participants are at a similar development stage, they have a lot to learn from...Sometimes it is helpful to find peers that share a similar direction or problem. For example, a group of people...most hits. Commit to a meeting schedule and meet regularly no matter what, even if one person can’t make ... work at the same place, a 1-2 hour lunch on a regular schedule might work. If not, perhaps a 3 hour breakfast... handy list of suggestions.  Get started. Have regular meetings. Experiment with the process. Have each...

  18. Antibodies 101: Fab Fragments

    Type
    Blog Post
    Published
    Sept. 5, 2024, 1:15 p.m.
    ...described above can be used for similar applications and have similar advantages, though with some caveats...chain. Created with biorender.com     The modularity of antibodies means that it is relatively easy..., two full antibodies bound together may be too large. Using a complex that is one full antibody bound...Alternatively, Fab fragments can be used as a block in similar scenarios. By applying unconjugated Fab fragments...and have lower binding affinity than some of the larger fragments.  Think of antibody fragments as new ...

  19. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ... we will focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB...start with a brief history of the gene and how molecular biologists have harnessed it to evolve cloning...clones.   ...Becomes a powerful cloning tool Molecular biologists first saw the potential of this system...and are considered F-); however, there are a few popular lab strains, such as NEB Stable, JM109, XL1 Blue...experiments. Please check out Addgene’s Choosing a Molecular Cloning Technique page and the Addgene Blog for...Cloning Posts Resources on Addgene.org Choose a Molecular Cloning Technique Find Plasmid Cloning Protocols...

  20. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    ...can be particularly useful in academic labs when creating shared experimental protocols. Clarify: Asking...setting, where student and postdoc turnover happens regularly and it behooves everyone in the lab to be using...Asking for clarification during a meeting can help get everyone on the same page. Others in the meeting might...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing 8-10 ...understanding you can be incredibly frustrating. Clarifying the words —> decreasing frustration —> increasing...
Showing: 241 - 260 of 811 results