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We narrowed to 1,005 results for: RING

Showing: 401 - 420 of 1005 results

  1. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...genome replication, repairing mismatched basepairs or small indels that occur during DNA synthesis, and...restriction enzymes are one half of naturally occurring restriction modification systems that prokaryotes...DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired mutation...methylation Finally, you can control methylation by altering your choice of bacteria. For example, if you must...

  2. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...most important step in ensuring quality, as it will catch any errors made during the cloning, sample handling...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...samples coming through our doors, the first step to ensuring plasmid quality is to stay organized. Here at ..., antibiotic resistance, restriction sites used during cloning, and most importantly, the predicted full...

  3. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ...retinal ganglion. Rather than using naturally occurring promoters to drive transgene expression, the Roska...one type of cell, with the remaining expression occurring in only Müller glia cells, a common contamination...retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic promoters performance in mouse, monkey...targeted cells in monkey and human retinas. By comparing the performance of promoters across these three...be paired for targeted expression. This same pairing of synthetic promoters were also used to target...

  4. A Look at Addgene's QC Process

    Type
    Blog Post
    ...completely different than what was originally intended. Sharing unverified plasmids can lead to far reaching consequences...standardization provides security for requestors, ensuring that the plasmid they ordered contains the correct...QC topics that may require additional attention during analysis. These include topics such as: locations... features. Most of these regions are identified during the QC process, following sequencing and assembly...30% of deposited plasmids! We are committed to ensuring that we maintain a high standard of sequence accuracy...

  5. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...made of protein-coding genes, and you might be wondering what the rest of our genome could possibly be ... modulate ncRNA expression without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR...high specificity, particularly in applications requiring smaller delivery systems (Zibitt et al. 2021).... a number of different options. By carefully considering your ncRNA of interest, you should be able to...RNA-targeting CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer...

  6. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ...their promises, Wet-blanket negativist, who can bring down the morale of a group with their pessimism,...communicate their ideas confidently while also considering the needs of other people. Some students and ...Regardless of your supervisor’s management style, bring challenges to his or her attention as soon as you...signed, or manuscripts that need to be reviewed, bring them to the meeting. This will make it easy for ...problem by stating the facts and explain how it is interfering with your work. Avoid talking about your emotions...

  7. Transferable Skills: Negotiation

    Type
    Blog Post
    ...something done not by people wearing a lab coat, but instead by people wearing business suits discussing ...transferable skills while working in the lab and by offering advice on areas where you might need some help...you are negotiating. Then take a moment before, during, or after a negotiation to think about the situation...experience in the area I want to explore while also ensuring that my work gets completed? I want my company...

  8. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...special considerations. Conventional flow cytometry During conventional flow cytometry, light emitted by a...distinguish these two fluorophores from one another during spectral unmixing.     Figure 2: The ...a three color panel or a 40 color panel! When preparing a spectral flow experiment and panel, there are...telling exactly how many Cy7 molecules are added during that particular batch. This can present particular...upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical to include...

  9. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...perform functional tests. The newest genome engineer sharing the stage with much-lauded CRISPR-Cas9 is DNA-guided... endonuclease NgAgo! We'll discuss how NgAgo is faring with users in a minute, but, to start, let's review...cleavage (1). Both the CRISPR-Cas9 and NgAgo genome engineering methods have their own pros and cons (2). The...protein to load an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo...Development and applications of CRISPR-Cas9 for genome engineering." Cell 157.6 (2014): 1262-1278. PubMed PMID: ...

  10. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...Introduction There are three primary methods for repairing DNA after a double-stranded break. HDR copies ...mutations seen with TALENs and CRISPR. MMEJ is active during M and early S phases, when HDR is inactive, and...Non-homologous End Joining Read About Other Genome Engineering Tools like Sleeping Beauty Mutase Find the Ideal... genome engineering... Mary Gearing...

  11. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...heterozygote into a homozygote and ensuring that all future offspring inherit the endonuclease gene. As...As we pointed out last year2, delivering a DNA cassette encoding the cas9 gene and single guide RNAs (...shared by everyone, he is deeply concerned with ensuring that gene drives and other collective technologies...selfish genes as tools for the control and genetic engineering of natural populations. Proc. Biol. Sci. 270,...

  12. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...presence of antigens (such as viral vectors) without triggering a destructive inflammatory immune response. At...after the cloning of the RPE65 gene, a naturally occurring animal model, the RPE65−/− dog, was characterized...retinal neurons. Nowadays a toolbox of naturally occurring and engineered AAV serotypes that further expands...being done to further improve the technology: engineering of new capsid variants, designing expression ... optimize transgene levels, and large scale manufacturing of viral vectors to support trials for prevalent...

  13. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ... the Addgene blog Learn more about plasmid sharing during COVID-19 Resources on Addgene.org Find plasmids...False, type='scriptV4', hide_playlist=True, viral_sharing=False, embed_button=False, autoplay=False, hidden_controls...to follow an exponential decay model, with dips during weekends and significant holidays. Figure...likelihood by the corresponding factor 𝜙z if t is during event z. The "events" we considered for our model...

  14. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    ...If you're interested in sharing your science, one of the big questions you'll be faced with is "how?"...for longer educational videos, and Twitter, for sharing updates, new deposits, and new blog posts!  ...  of proficiency.  What kind of content are you sharing?  Sometimes your subject matter will drive the ...fair, or a semester-long after school program? Comparing your desired outcome to the limitations and potential...response. High connection levels and interaction. Can bring in things to be physically interacted with. Public...

  15. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses...producer cell membrane, and acquired as an envelope during exocytotic budding, as the viral particle emerges...transfer of RT, which would actually occur in step 2 during uncoating. After uncoating, reverse transcription... integrated by the viral integrase protein (IN) during step 3 (King et al., 2008) (Hu & Temin, 1990). ...

  16. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...suppose you have a piece string that’s 10 meters long. You twist and roll the string into a tight ball. Now...in a way that brings linearly distant regions close together. gRNA Selection and Filtering for Off-target...Now the two ends of the string, instead of being 10 meters apart, are mere millimeters apart. Similarly...

  17. CRISPR Antimicrobials

    Type
    Blog Post
    ...bacteria and prevents sensitive bacteria from acquiring resistance genes. Yosef et al. suggest that it...not previously infected by the lysogenic phage, rendering the population sensitive to antibiotics. Created... Lemire, Diana P. Pires, and Timothy K. Lu. “Engineering Modular Viral Scaffolds for Targeted Bacterial...Addgene Blog CRISPR Methods for Bacterial Genome Engineering Treating Muscular Dystrophy with CRISPR Gene ... Mary Gearing...

  18. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...viral genome). Empty viral capsids can be formed during viral production depending on the  experimental...usually report? Addgene and other viral vector manufacturing facilities report the physical titer of a viral...to two-fold. How can we get more reliable AAV titering? For our viral service, we currently titrate AAV...different sources in the same experiment, consider retitering both AAV in your lab. The absolute titer values...Addgene, we don’t reserve lots, but if you’re reordering the same virus feel free to email us and we can...

  19. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ... data. Although it may be useful to selectively bring particular items into an ELN – e.g. an image that...the integration with protocols.io, the protocol sharing repository. As illustrated in this video the integration...RSpace becomes part of a workflow that enhances the sharing and reproducibility ecosystem protocols.io makes...help with searchability, record-keeping, and data sharing, not all of them support reproducibility in the...Reproducibility Read all blog posts about scientific sharing Resources on Addgene.org Find protocols of molecular...

  20. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...editing reporters that can help you do just that. Monitoring APOBEC and Cas9-mediated editing in real time...results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using...editing efficiency can be easily determined by comparing cells expressing only eGFP expression to cells...real-time, it relies on DSBs which is not ideal for monitoring single base edits (St. Martin et al., 2018). ...the Harris Lab circumvents the need for DSBs in monitoring on-target DNA editing efficiency of APOBEC-Cas9...
Showing: 401 - 420 of 1005 results