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We narrowed to 922 results for: POR

Showing: 541 - 570 of 922 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...transduced rat neurons in vivo with beta-gal and other reporters, and they showed that expression persisted for...Figure 2). The HIV genes that do remain are very important for viral production: Gag (structural precursor...activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A different viral envelope protein...

  2. CRISPR Antimicrobials

    Type
    Blog Post
    ...species, or even an antibiotic resistance gene. Importantly, native, non-pathogenic bacteria remain to help...recolonize the niche, reducing the chance of an opportunistic infection like for organisms like C. difficile...colonization model, phagemid treatment decreased the proportion of virulent S. aureus from 50% to 11% in just...

  3. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...After sequencing and searching for cellular RNAs important for Cpf1 function, they found that mature crRNAs...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions...

  4. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...RNA editing - and further experiments strongly supported their initial findings.  The team originally suspected.... Specifically, efficacy rates of Cas7-11 were reported as up to 85%, broadly similar to shRNA and Cas13...However, in those same comparisons, the AbuGoot lab reported no detectable effect on cell viability – meaning...

  5. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...recent lineage tracing approaches that led to important insights in developmental biology. Microscopy-...pathway regulated by one-eyed pinhead, incorrect proportions of otherwise normal cell types are produced, ...PMCID: PMC6247916. Felker, A., and C. Mosimann. "Contemporary zebrafish transgenesis with Tol2 and application...

  6. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    .... More importantly, the expression of this construct is Cre-dependent, enabling spatiotemporal control...astrocytes, and oligodendrocytes, provided that reporter genes specific to these cell types are used. The...

  7. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...from 2016–2023 by type of tool. Equally as important to what is being delivered is where it’s being...vectors not only available, but accessible. The supporting data and community engagement required to do ...lessons learned from this process have shaped and supported our growth and development. We’ve shared what ...

  8. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in the pool...activity of Cas13a becomes activated and the RNA reporter will be cleaved resulting in activation of the...in the field and as always, we appreciate the opportunity to share these invaluable reagents with the scientific...

  9. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...this post will, indeed, focus on bacteria, it is important to note that the term ‘antibiotic’ can also be...animal farming is already being recognized as an important step, as you can’t walk into any supermarket now...Tackling drug-resistant infections globally: final report and recommendations." London: Wellcome Trust & ...

  10. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...Labor Thomas Perez, Fair Pay for Postdocs: Why We Support New Federal Overtime Rules, made clear that institutional...National Postdoctoral Association Institutional Policy Report 2014). However, ~30% set their minima lower. Furthermore...consensus of academic stakeholders. There was ample opportunity to accept these recommendations, and make gradual...

  11. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...2016; Lambert, 2019). These characteristics are important, but have less priority than stickiness. In my...lead to over-expression artifacts. This is an important downside of the technology and should always be...fluorescent proteins in budding yeast." Scientific reports 9.1 (2019): 2234. PubMed PMID: 30783202. PubMed...

  12. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...split aptamer is composed of a “Top” and “Bottom” portion, each fused to a separate piece of RNA. Fluorescence...photostable, and were presented in the study to support the uniqueness of the photostability of the Corn-DFHO...growth as RNA biology, we are looking forward to reports on the next additions to aptamer soup! References...

  13. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...sequences, which facilitate integration of this portion into the host genome. The packaging plasmid contains...more detailed description). Since the integrated portion does not contain any of the viral packaging genes...the resulting double-stranded DNA transgene is incorporated into a large complex of both viral and cellular...

  14. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...signals. For this, you’ll need to gate based on the proportion between H and A (or W and A) within the same ... just as fine (Figure 4). As H and A scale in proportion (while H and W do not; see Figure 2), single ...                                                                                                         B.    Figure 4: (A) Using the proportional relationship between FSC-H and FSC-A we can ...

  15. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ...in competition with other similar integrins, as reported by IPI and the Springer lab (Hao et al., 2024)...In this study, we, along with the Springer lab, reported the binding specificity, kinetics, and affinity...belief that αVβ3 would be the most functionally important integrin on these cells. What’s next IPI is working...

  16. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...invasion and D loop formation, the newly synthesized portion of the invasive strand is displaced from the template...DSB-induced genome engineering, it has biological importance for the repair of breaks that lack a second end...Cut-to-Mutation distance" affects editing efficiency. One important point to note is that Cas enzymes can continue...

  18. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...mycoplasma contamination The best and the most important ways to avoid mycoplasma are by providing proper...viability. Conclusion To sum up, it is always important to prevent contamination rather than eradicating...others when you can and help them understand the importance of preventing contamination. Be aware and cautious...

  19. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...fibrillary acidic protein (GFAP) for astrocytes it is important to test it in the brain region of interest, especially...of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse... the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When Using AAVs Three Key Considerations...

  20. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...of the gene with multiple gRNAs, but the most important is that you could avoid potential off target artifacts...you could run another gel that highlights the important results (for example, if you needed to make a ... Despite the shifting landscape of research opportunities, it’s an exciting time to be in the lab. Based...

  22. CRISPR Library Amplification

    Type
    Protocol
    ... mix. Electroporate cells (one at a time for a total of eight electroporations): Electroporator Conditions...and have 1 mL SOC per electroporation readily available for post-electroporation recovery of cells. Ensure...if the recombinant band makes up a significant proportion of the DNA pool. Last Update: August 17, 2023...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents.../or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad, Micropulser, 0.1 cm ) 20 ...convenience but any brand or house- made LB that supports normal growth is expected to work. 50 mL Falcon...DNA-Endura into pre-chilled cuvette. Pulse. Caution Electroporation involves the use of high voltages, please use...

  23. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ... DNA to favor the edit Once the prime editor incorporates the edit into one strand, there’s a mismatch...MutSα–MutLα mismatch repair (MMR) complex, to temporarily inhibit this process and promote copying of the...it from hybridizing with the unedited strand. Temporarily inhibiting this complex may allow time for 5′...

  25. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    .... They also mutagenized the cytidine deaminase portion of the base editor to create SpCas9 base editors...off-target activity (Richter et al., 2020). This is important because ABEs have generally been slow, meaning...editing for plants Read about four base editor reporters Additional Resources on Addgene.org Catch up...

  26. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...Product Run PCR to amplify your insert DNA. It is important to use a high fidelity taq polymerase to minimize.... The fidelity of the polymerase becomes more important the longer the expected PCR product is. You should...temperature based on the melting temperature (Tm) of the portion of the primer that hybridizes to the sequence to...some DNA during the gel purification step, it is important to digest plenty of starting material. We recommend...be cut with both enzymes, and therefore it is important that the digest goes at least 4 hours and as long...running a gel for purification purposes it is important to have nice crisp bands and to have space to ...expected. When cloning by PCR, it is especially important to run the product on a gel. This allows you to...

  27. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...446 Benzamide, Millipore Sigma 12072 Antipain, Millipore Sigma 10791 Leupeptin, Millipore Sigma L2884 Aprotinin...monobasic monohydrate, Millipore Sigma 71507-250G Sodium phosphate dibasic, Millipore Sigma S7907-500G 1 ...that sharing the full details of our protocols supports reproducibility and accelerates science. Here,...information is solely for transparency intended to support reproducibility in science. Safety Warnings Sodium...1 M Trizma hydrochloride solution pH 9.0, Millipore Sigma T2819-1L 250 mL sterile bottle, VWR 430281 0.45... Aprotinin, Millipore Sigma A6279 Zeba Spin Desalting Columns, 7 kDa MWCO, 10 mL, Thermo Fisher 89893 ...89893 Amicon Ultra-15 30 kDa MWCO column, Millipore Sigma UFC903024 50 mL LoBind tubes, VWR 76289-498 Before...

  28. Trends in CRISPR and SynBio Technologies

    Type
    Blog Post
    ...reflections on the program are here. This was a great opportunity for Addgene to present our own data on plasmid...in advancing this field and how we are able to support rapid progress in all areas of research. We welcome...

  29. New FLIS Plasmid (April Fools 2022)

    Type
    Blog Post
    ...more commonly known as fruit flies, have been an important animal model for research, particularly genetics...The Frequency Light Interval System, or FLIS, incorporates a bioluminescence protein into the chitin that...

  30. Twenty Years of Sharing Science!

    Type
    Blog Post
    ...India, and Brazil.  We've also created a robust support system for scientists using materials in our repository...repository, from our customer and science support to our efficient and easy tech transfer process to our...
Showing: 541 - 570 of 922 results