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We narrowed to 932 results for: omp

Showing: 341 - 360 of 932 results

  1. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...specific chromosome. This image represents the composite of fluorescent measurements from red, blue, and...modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly, thereby decreasing the amount of background... labeling efficiency was similar to that of a comparable FISH method. Importantly, they did not observe...with the previously established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that...that they could evaluate the degree of chromatin compaction even for this small distance! To the authors’...CRISPRainbow generates 6 unique colors and a white composite. gRNAs with two identical hairpin binding domains...

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

    Type
    Blog Post
    ...specific components that are key to the cloning reaction. Understanding the specific components is essential...take up your insert. But exactly how does ccdB accomplish this? Let’s start with a brief history of the...system contains ccdB. A successful insertion will completely replace ccdB with the investigator’s insert of...interest.   CcdB resistant E. coli strains complete the system But ccdB alone isn’t the only key to...protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3):735-45. PubMed...

  3. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...was contributed by guest blogger Alan Wong. The complexity of biological systems can hinder our attempts...simple tool that allowed us to rapidly decode the complexity? The CombiGEM-CRISPR technology was developed...of providing an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your ...mapping the combinatorial genetics that underlie complex regulation in biological systems. The CombiGEM ...new technologies to interrogate and understand complex biological systems. He can be contacted at aslw...transcriptional activation by an engineered CRISPR-Cas9 complex." Nature (2014). PubMed PMID: 25494202. PubMed ...

  4. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ... do as genetic circuits become larger and more complex? Voigt: If you look at all the products of biology... on genes at different times. It requires more complex synthetic networks. Addgene: What hurdles did you... perspective and you can layer them for more complicated functions. Those things are easy with CRISPR..... There haven’t been enough genetic parts for complexity. We have characterized almost 600 terminators...of a cell. The objective is to give people the complete toolbox for taking on very large genetic engineering...just using what biology has given us. Wood is a complicated material and we can access it, but we are stuck...

  5. A Look at Addgene's QC Process

    Type
    Blog Post
    ... to stop functioning properly or be something completely different than what was originally intended. ...tags, promoters, and other critical functional components. Fun fact! Some older plasmids in our collection...each of their plasmids, from descriptions of key components of the plasmids to partial, full, or annotated...Addgene receives deposited plasmids, they undergo complete plasmid sequencing using an in-house Illumina ...Addgene. The assembled plasmid sequence is then compared to the information provided by the depositor. ... sequence is not available, QC scientists will compare the NGS results to the plasmid information, other...

  6. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...that allow their host organisms to thrive in competitive environments. The genes encoding these functions...triumph over your neighbors is to eliminate the competition, and Penicillium isn’t the only organism that... are also less aggressive ways to survive in competitive environments — like by living where no one else...to be active only in the presence of the target compound. These systems are a gold mine of parts for synthetic...Shareck et al., 2004). Plasmid categorization is complex Plasmid classification has shifted over the years...necessarily reflect genetic relationships or plasmid incompatibility. Plasmids may also have more than one function...

  7. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...science and collaboration. The field already has comprehensive reference pages and curated databases for scientists... (CeNGEN) The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron...that is creating the “first virtual organism in a computer.” This project has produced various resources ...four-dimensional contrast interference (DIC) microscopy and computer imaging processing. The combination of these methods...methods provides unique opportunities to develop computational methods to better understand animal development...Hammarlund, Marc, et al. "The CeNGEN Project: The Complete Gene Expression Map of an Entire Nervous System...

  8. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...blood clotting, and as we all know, a structural component of bones and teeth. It is an enzyme co-factor,..., a neuron subtype) and specific subcellular compartments for measurements. All of the sensors discussed...calcium concentrations in different subcellular compartments can vary greatly. Additionally, the calcium ...susceptible to artifacts and pH interference, and comparison between experiments can be more difficult due...GCaMP6 only) or mammalian cell expression, and compatible with constitutive or Cre-dependent expression...Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo. Neuroscience Research...

  9. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...significant amount of flexibility compared to a strict single-compatibility sequence like NGG. Nonetheless...NG, a rationally engineered Cas9 with NG PAM compatibility, as the name implies (Nishimasu et. al, 2018...know and love about Cas9, but with broader PAM compatibility. This alternative PAM requirement adds a significant...PAMs and is capable of editing at frequencies comparable to SpCas9. SpRY can also edit NYN PAMs, but with... Ai C, Hu J. In-depth assessment of the PAM compatibility and editing activities of Cas9 variants. Nucleic...

  10. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...remodeling. Chromatin is a complex of DNA and histone proteins that compacts cellular DNA into chromosomes...in the compacted region and results in a loss of gene expression. In this way, genes in compacted regions...and tightly bound DNA is to histones, the more compacted it becomes. This prevents the cellular machinery...have huge impacts in the way we study and treat complex or multifactorial diseases, enabling faster diagnosis...

  11. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...information directly into the file. Email the completed spreadsheet to [email protected] along with ...used to knockdown expression of mRNAs with complementarity to the shRNA sequence. Empty backbone A plasmid...plasmid data in the pages described below. You can complete these pages sequentially by entering the appropriate...the links at the top of the page. Figure 11: A completed plasmid page for review before submission. Once... right of each data entry page. Once you have completed the data entry process for all of the plasmids...Figure 13: Click the "Submit Deposit" button to complete the data entry. Clicking the "Submit Deposit" ...third party rights holders. Once this process is complete, you will receive a Deposit Confirmation email...

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

    Type
    Blog Post
    ...bacterial strains, like NEB Stable. NEB Stable competent cells lack RecA, a protein that aids in the homologous...helps me figure out if I have enough plasmid to complete my experiment. AAV is usually the limiting reagent...produce more virus because I don’t have enough AAV. Compared to the Addgene AAV Production protocol, I take...concentrate the purified virus. This process can be completed in one long day, or the virus can be stored at...set can be used and samples are quantified by comparing to an AAV standard curve, which is generated from... plasmid. This whole process takes ~3 hours to complete: 1 hours hands-on time and 2 hours for the qPCR...

  13. PhD Applications After COVID

    Type
    Blog Post
    ...know that this process only became even more complicated with COVID-19 and standards for hybrid, virtual...without the financial and logistical stress that accompanies traveling for campus visits. This is definitely...where the PhD application cycle becomes much more complex. “How do I reach out to potential mentors?” “Where...very valid questions, and often make an already complicated process even more difficult. I’ve learned that...to be incredibly useful, as it gives a more comprehensive idea of that advisor’s research interests which... for the interview. Having notes close to your computer with things you want to highlight in the interview...

  14. Transferable Skills: Negotiation

    Type
    Blog Post
    ...while also ensuring that my work gets completed? I want my company to pay for me to attend a conference... How can I negotiate my starting date with the company so they know I’m excited while still being able...want to learn more about a different area of the company. How can I negotiate with my manager to gain experience...conference. How can I negotiate to show the company that investing in me is also beneficial to them? I have an...but I need help from another person/team at the company. How do I negotiate with the person/team for help...

  15. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...about a month and a half, without the need for complicated cloning of homology arms. MMEJ: An Introduction...HDR and NHEJ, please see the linked blog posts. Compared to its counterparts NHEJ and HDR, MMEJ doesn’t...strategy looks very similar to HDR, but it is accomplished using much smaller regions of homology, which...strategy looks very similar to HDR, but it is completed using much shorter sequence homologies. Abbreviated...efficiency of PITCh-based genomic insertion would compare to CRISPR sticky-end insertion using the nuclease...Genome-Editing Based Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: ...

  16. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...These insights are hard to appreciate without computational support. Indeed, rational sgRNA design is not...experimental applications in CRISPR engineering. When compared to that collective utility, it’s easy to fault...tool unique. The CRISPR Software Matchmaker is composed of these features and enables you to select the... based on their comfort level with operating a computer.  Examples: “offline”, “online”, “CLI”, and “GUI...few offline tools available mostly work on any computer but generally require working knowledge of the... power of each tool they are interested in by comparing their past results to the designs suggested by...pattern at any edit distance. It won’t, however, compute any score other than the edit distance. Added functionality...

  17. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...just one or two recombination events, homemade competent cells of 107 CFU/μg are perfectly fine, yet performing...performing multiple modifications will require ultracompetent (109 CFU/μg) cells. Primer design Using specific...insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer design for each...the forward primer by adding a short sequence complementary to the 5’ end of the reverse primer. Mutagenesis...efficiency of assembly decreases as the cloning complexity increases. The efficiency of recombination is...increased with greater homology, so performing more complex procedures can be aided by extending homologous...

  18. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ...Sharing data and community concerns AAV vectors are complex biological reagents; many factors, like storage...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data...through word of mouth or old lab notebooks and computer files. If you asked your team members or colleagues...much unpublished AAV data do you have sitting in computers and notebooks right now?, I’m willing to bet the... rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https..., C., & Flotte, T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under ...

  19. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...division. These non-plasmid-containing bacteria can compete with and confound the goal of your experiment —...spectinomycin is the more stable and expensive antibiotic compared to streptomycin. Spectinomycin/streptomycin can...option! Otherwise, since it isn’t universally compatible with all bacteria used in research, it isn’t ...express the Tn5 element, but thankfully, most competent cells do not express this factor. Zeocin is handy...host/plasmid DNA during selection if it’s not completely bound up. While this likely occurs at a low rate...Stability Time Saving?  Works for multiple organisms? Compatible with most bacteria? No satellite colonies? ...

  20. An Introduction to Adenovirus

    Type
    Blog Post
    ...immune system; and reconfiguring of the nuclear compartment, gene expression, and protein translation. Its...producer cells where there is E1 and E3 gene complementation. In target cells that are infected by the vector...virions) are not expressed. Note that replication-competent AdV vectors also exist and are used in certain...topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, and adenoviral...higher than other viral vectors (tens of kilobases; compared to only 4.7 kb for AAV, 9 kb for retroviruses)...Tales: From the Cell Surface to the Nuclear Pore Complex. PLoS Pathog, 11(6), e1004821. https://doi.org/...
Showing: 341 - 360 of 932 results