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We narrowed to 328 results for: hal.1

Showing: 181 - 200 of 328 results

  1. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...for example, split cells on a Monday diluting them 1:10, you should be able to split on the following Thursday...Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis...Turning on the UV light 3-4 times a week for up to half an hour can help keep your hood contamination free...Additional Resources on the Addgene Blog The Challenges of Cell Culture Cloning Mammalian Cells with ...

  2. QC Sequencing Technologies at Addgene

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    Blog Post
    ...and recorded as a DNA strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions...trace” file to determine the sequence. Figure 1: Overview of Sanger sequencing technology. Created...Technologies. Current Protocols in Molecular Biology, 122(1). https://doi.org/10.1002/cpmb.59  Wang, Y., Zhao,... up the pace Our QC team constantly faces the challenge of keeping up with the large amount of QC analysis...

  3. Hot Plasmids: Fall 2024

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    Blog Post
    ...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9... Zhong, B. L., Elliot, J. M., Wang, P., Li, H., Hall, R. N., Wang, B., Prakash, M., & Dunn, A. R. (2024...

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

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    Blog Post
    ...catalog had grown to 879 viral catalog items (Figure 1). The service caught on quickly in the scientific ...over 100,000 viral vector preps!   Figure 1: A) Cumulative AAV preps available in the Addgene ...Neuromethods, 195, 77–101. https://doi.org/10.1007/978-1-0716-2918-5_5 Haery, L., Deverman, B. E., Matho, K...depending on their serotype, are particularly challenging. Their efficacy in any given system can be affected...

  5. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    Blog Post
    ...guide RNA (sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the...bystander bases” if you choose this system.  Fig. 1: Cytidine deamination takes place on the free strand...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...essential genes whose knockout would cause cell lethality. The Krüppel associated box (KRAB) is a naturally...

  6. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ...In vitro screening for the best gRNA Figure 1: Validation of double strand break (DSB) mediated ...as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more...and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target effects...stranded break at the target site, allowing the two halves of EGFP to recombine by homology directed repair...

  7. Important Considerations in Optogenetics Behavioral Experiments

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    Blog Post
    ...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...follow him on Twitter @derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission...than ICSS. The test chamber is divided into two halves that are distinguished visually through different...

  8. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...replication and also act as signals for packaging. Figure 1: Wild-type AAV genome. Created with BioRender.com....extensively studied, and has a broad tissue tropism. Table 1 gives a summary of the tropism of AAV serotypes, indicating...AAV5, AAV8 Skeletal muscle AAV1, AAV8, AAV9 Table 1: Summary of tissue tropism displayed by different ...transgene, but also at a very low efficiency (less than 1% of wild-type). Another split AAV method which has...vectors pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell...short coding sequences of channelrhodopsins, halorhodopsins, and other optogenetic genes makes them suitable...) and a gRNA together into an rAAV capsid is challenging due to its limited packaging capacity, especially...

  9. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene...the years. The authors optimized these systems by: 1) decreasing the number of plasmids required for each...

  10. Your Top Requested Plasmid in 2015!

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    Blog Post
    ... another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...only been available for a little over a year and a half, a small sampling of the research it has already...PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide...

  11. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...oligodeoxynucleotides)) are commonly used for smaller modifications (~1-50 base pairs). Small edits require as little as 30...Easi-CRISPR was shown to increasing editing efficiency from 1-10% with dsDNA to 25-50% with ssODNs in mouse models...ribonucleoproteins. Quadros RM, et al. Genome Biol. 18(1); (2017). PubMed.  Additional Resources on the Addgene...CRISPR forum moderated by the Zhang lab CRISPR Challenges: Standardization and HDR CrispyCrunch Optimizing...

  12. Antibodies 101: Flow Cytometry

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    Blog Post
    ...interrogated by the optical system (Figure 1).   Figure 1: Cells expressing a particular surface ...populations of cells in a complex tissue or sample (1). When combined with cell sorters, this technology...then flow cytometry can help you overcome these challenges and make these types of experiments easier and...fluorophores Fluorescent antibodies get around the challenges of fluorescent protein fusions by binding to ...

  13. Predicting Adverse Reactions to Monoclonal Antibody Drugs

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    Blog Post
    ...immunogenicity score (Mattei et.al., 2022).    Figure 1: Generation of immunogenicity scores by predictive...antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020). https://doi.org/10.1186/s12929-019-0592-z  ...Formation in Oncology: Clinical Relevance and Challenges. The oncologist, 21(10), 1260–1268. https://doi.org...

  14. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

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    Blog Post
    ...prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who believe...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the...lockdown forced us to put our research projects to a halt. None of us works directly with viruses or transmissible...

  15. Viral Vectors 101: The AAV Retrograde Serotype

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    Blog Post
    ... There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to...significantly updated in 2023 by Jason Nasse.  References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant...recombinase-dependent vectors, this can cause challenges for the experimental design as these vectors ...

  16. Plasmids 101: Protein tags

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    Blog Post
    ...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function... conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based ...purify proteins can face any number of technical challenges depending on their protein of interest. After...After several decades of trying to address these challenges, researchers have amassed a considerable molecular...

  17. Antibodies 101: The Four ELISAs and When to Use Them

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    Blog Post
    ...alternative detection methods.      Figure 1: The four types of ELISA are direct, indirect, sandwich...conjugated antibody is not readily available. Table 1: Overview of the advantages and disadvantages of each...antibodies. Assay development, however, can be challenging since scientists must first identify two independent...Longer protocolIncreased chance of cross-reactivityChallenging to develop assay Competitive Appropriate...

  18. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ... medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our research is to enhance ...activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong...transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant can take...Luciferases usually have shorter mRNA and protein half-lives then GUS and are therefore good temporal reporters...

  19. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...and drawbacks of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy has...Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27:1–12 . https://doi.org/10.1089/hgtb.2015.140 Frischmeyer...packaging capacity is limited to ~4.7kb, or roughly half the packaging limits of lentiviral and adenoviral...multiple copies of both vectors. Examples of use: Halbert et al., 2002 used an overlapping vector with 440...heterodimer, then splicing of the pre-mRNA of the two halves of the transgene will reconstitute the full-length...A of the transgene contains the promoter, the 5’ half of the transgene, a splice donor sequence and the... of homology, a splice acceptor sequence, the 3’ half of the transgene. The full length gene is reconstituted...

  20. Our Most Popular Blog Posts from 2019

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    Blog Post
    ...deletions, and all possible base-to-base changes.   1. Plasmids 101: Transformation, Transduction, Bacterial...and represent the diversity of tools we share on behalf of scientists. 9. Plasmids 101: Positive and Negative...fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview by Erik Snapp...blogger Erik Snapp breaks down how to prepare a chalk talk for an academic job interview including defining... research vision, organizing your vision into a chalk talk, and tips for practicing.  6. The Fluorescent...
Showing: 181 - 200 of 328 results