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We narrowed to 672 results for: abo.1

Showing: 241 - 260 of 672 results

  1. Special Delivery: Fluorophore Targeting for FRET Studies

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    Blog Post
    ...standard for genetically-encoded FRET donors (Fig. 1). Scores of FP variants have been created, thereby...with finesse using tRNA suppression technology (Fig. 1) (Dumas, Lercher, Spicer, & Davis, 2015). By using...orthogonal labeling strategies are presented (Fig. 1) which can be carried out using Addgene plasmids. ...involved in skeletal muscle contraction.   References 1. Degorce, François, et al. "HTRF: a technology tailored... transfer measurements of ryanodine receptor type 1 structure using a novel site-specific labeling method...environment? If these are pressing issues in your laboratory, then FRET experiments are a viable biophysical...to consider these cases separately when talking about the tools that are available. Let’s consider the...

  2. Grad School Advice Part 1: Picking a Lab and a Project

    Type
    Blog Post
    ...Ben describe their projects 0:28 - 1:12 : Introduction from Tyler 1:13 - 2:02 : Niroshi and Ben introduce...graduate school community to help keep you excited about coming into lab even when things get tough in the...

  3. RANbodies: Reporter Nanobody Fusions

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    Blog Post
    ...applications. Read on to learn more about each reporter or check out table 1 for a summary. HRP HRP tagged ...with a standard primary/secondary antibody. Table 1: Summary of the Key Features of HRP, Spaghetti Monster...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...source of monoclonal antibodies, but they are laborious to generate and costly to share. What is a RANbody...Additional Resources on the Addgene Blog Learn more about using the secondary nanobody toolbox for immunodetection...can be found here Watch this video to learn more about nanobodies Additional Resources Looking for primary...

  4. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ... employing HDR. Ma et al. moved forward with gRNA 1 to conduct microinjection experiments in S phase zygotes...instead from the mother’s wild type allele (Figure 1). 72.4% of embryos carried two wild-type copies of...proceeded through NHEJ rather than HDR. Figure 1: Homology-directed repair in targeted zygotes proceeds...August 2nd in Nature gives us even more to talk about, as it represents another step towards CRISPR germline...impossible with current tools, thus quieting some fears about the creation of “designer babies.” The differences...of individuals with varied backgrounds. Talking about this study with non-biologist friends Like me, you... Blog Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes...

  5. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

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    Blog Post
    ...neuron which generates an action potential (Figure 1). This light-induced excitation can take place in ...channelrhodopsin activation to a small volume of brain tissue (~1 mm3). Current channelrhodopsins also need high-intensity...fiber-optic cables into the brain. Figure 1: With no blue light, the channelrhodopsin remains ... blog Read more about the PHP.eB AAV capsid Read more about optogenetics Learn about the differences ...library were designed using structural information about parental channelrhodopsins to create recombinant...

  6. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...dual-color control of neurons. Nature Communications, 12(1), 1–20. https://doi.org/10.1038/s41467-021-24759-5 Zhang...to consider when choosing one.     Figure 1: Example of an experimental optogenetic setup. A light... 21–51). Springer US. https://doi.org/10.1007/978-1-0716-0755-8_2 Lehtinen, K., Nokia, M. S., & Takala... 21–51). Springer US. https://doi.org/10.1007/978-1-0716-0755-8_2 Boyden, E. S., Zhang, F., Bamberg, E...which tools to use, and what do you need to know about them to be successful in your experiments? In this...

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

    Type
    Blog Post
    ...effective recovery of knock-ins, but did result in 1-2 kilobases of additional sequence being introduced...facilitating recovery of edited animals. Figure 1: In the background are 504 restriction digests of ...selection for repair of a temperature-sensitive pha-1 mutation significantly enriches for knock-in of 2x... 3xFLAG epitopes into other, non-linked loci; pha-1(e2123) mutant worms are perfectly viable at 15 ºC,... repair pathway. Reagents required to perform pha-1 co-conversion are available through Addgene. In 2019...learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about gRNA design...interest, I encountered low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering...

  8. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ...the other (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv. ...intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged with 10-24 copies...Application. Clinical and Developmental Immunology 2012:1–15 . https://doi.org/10.1155/2012/980250  Bates A,...without disulfide bonds, made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology... Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more about antibodies...Köhler who developed hybridoma technology. But what about single chain fragment variables (scFvs)? scFvs weren...from inclusion bodies, it’s time-consuming and laborious.  To avoid this, one or more of these approaches...

  9. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

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    Blog Post
    ...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...promote scientific sharing. Listen to learn more about Darrell, his research, and his focus on open science...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Podcast.     Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids...Plasmids from the Kotton Lab Learn more about Stem Cells ...

  10. RNA Extraction Without A Kit

    Type
    Blog Post
    ...glassware should be treated with a DEPC solution for 1 hour, and autoclaved to remove residual DEPC. Alternatively...more large, expensive ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction...extraction: twenty-something years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot.2006.83 Additional...more foolproof (still without a kit!) As mentioned above, working with RNA requires keeping your samples ...lysis. This can be challenging depending on your laboratory situation or tissue collection method, so biotechnology...supernatant is extracted with chloroform as in the above no-kit method.  Researchers have also developed ...

  11. Antibodies 101: Conjugation

    Type
    Blog Post
    ...domain (Figure 1).  Figure 1: Examples of sites on antibodies used for conjugation. 1) Endogenous ...Check out our post on Biotinylation to learn more about biotin and its use in molecular biology. Other Conjugates...perfectly blissful, productive lives without thinking about these chemical reactions. After all, there is an...ready to use off the shelf. So why are we talking about this? Even though there are so many commercially...

  12. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ...separation is maintained by 2 sets of interactions : 1. Interactions between the non-target strand and a ...theoretically decrease off-target cutting. Figure 1: Strand separation and target DNA binding to the Cas9... improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered ...With this new found structural knowledge, in collaboration with the Joung Lab, they used targeted mutagenesis...edits in mammalian cells and may decrease worries about off target effects in applied and/or therapeutic...at Addgene. Resources on the Addgene Blog Read about Cas9 Homolog Cpf1 Read How Non-Homologous End Joining...

  13. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...GFP expression.  Cloning and expressing RNAi pMKO.1 puro (Plasmid #8452) RNA interference (RNAi) is an...information and a detailed protocol on how to use the pLKO.1 vector can be found here. Cre-Lox technology Puro....of their choice in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi! Cre-Lox technology...knockdown. Vector provided by the Qi lab. Learn more about CRISPR plasmids for transcriptional inhibition. .... Check our CRISPR guide and blog to learn more about CRISPR and the most recent developments in the genome...

  14. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

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    Blog Post
    ... the samples stored in our lentivirus tubes (Fig. 1). However, once thawed, there was a substantial reduction...in the pH of samples stored in our AAV tubes (Fig. 1). It’s important to note that the change in pH only...using the thawed virus within 24 hours. References 1. Murphy BM, Swarts S, Mueller BM, van der Geer P, ...return the sample back to the original pH. What about putting it back in the freezer? We then tested whether...

  15. Plasmids 101: Biotinylation

    Type
    Blog Post
    ... of the biotin-avidin interaction   Figure 1: Biotin is enriched in egg yolk while its binding ... substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular Biology...matrices.” Journal of Nanobiotechnology 13.1 (2015): 1–14. PubMed PMID: 25980573. PubMed Central PMCID: PMC4461904...detection by northern blotting.” Rice 7.1 (2014): 1–7. PubMed PMID: 26224555. PubMed Central PMCID: PMC4884002...uses of biotinylation in molecular biology! Learn about in vivo biotinylation of bacterial fusion proteins...covalently attached to the active site of key metabolic enzymes including biotin carboxylase and decarboxylases... organic acids to produce important cellular metabolites. Biotin acts as a carboxyl carrier from one compound...

  16. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...s pLKO.1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...experiments.  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David ...collection Find plasmids for your research  Learn about our viral service ...

  17. Hot Plasmids - February 2022

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    Blog Post
    ...include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody (FB) construction from ...Comm. 2019. https://doi.org/10.1038/s41467-019-10846-1.   Optimized genetic screening technique using Cas12a... to Cas9’s larger expression requirements (Figure 1), but Cas12a has generally been underutilized in genetic... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  ...

  18. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ... requires selection. These two steps are (Figure 1): The initial plasmid integration step (the so-called...antibiotic resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads... loss of a specific phenotype (using sacB, Figure 1, further discussed below).  Counter-selectable marker...Current protocols in molecular biology 106.1 (2014): 1-16. PubMed PMID: 24733238. Additional resources on... a sucrase that converts sucrose into a toxic metabolite. Cells that have undergone a double crossover...confirmation of successful single crossover – after about 48 hours, the resulting colonies turn subtly blue...blog posts Find our CRISPR blog posts Learn more about Lambda Red recombineering Resources on Addgene....

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

    Type
    Blog Post
    ...succumb to the toxicity of CcdB.   Figure 1: Image from Bernard, P., et al. showing the pKIL 18...submit a guest blog entry of your own! References 1. Bernard, P. “Positive Selection of Recombinant DNA... plasmid ccdB killer gene”. Gene. 1994 Oct 11;148(1):71-4. PubMed PMID: 7926841. 4. Bernard, P., et al...in modern cloning, and why should you learn more about it? Read on to find out how ccdB can make your cloning...of this system for enhancing cloning efficiency about 20 years ago and developed cloning vectors to harness...

  20. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...were found on the plasma membrane (top panel in Fig. 1). Of course, this was problematic, because the DREADD...plasma membranes in both species (bottom panel in Fig. 1). Our results indicate that the tag protein may have...protein of interest the membrane.   Figure 1: Electron microscope images showing ultrastructural...Fluorescent 101 blog posts Read about chemogenetics vs. optogenetics Find more about the many types of protein...
Showing: 241 - 260 of 672 results