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We narrowed to 815 results for: tes

Showing: 261 - 280 of 815 results

  1. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...A4 and A8-A10. The researchers selected 8 of the tested ABE8 constructs (ABE8.8-m/d, ABE8.13-m/d, ABE8.17...2 guanines). This limits the use of ABEs as DNA sites that do not contain a suitable PAM cannot be edited...for primary human cell editing, the researchers tested ABE8 in CD34+ hematopoietic stem cells. ABE8 efficiently... gRNA used (Gaudelli et al., 2017). The authors tested ABE8.17-m and found that one base change mutation...overall improved base editing capacity, even at sites that used to be difficult to target. In particular...

  2. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...They will come in handy here! Crossing your F1 heterozygotes to each other will generate approximately 1 ...wild type at one or both of the alleles, or heterozygotes, are not a waste. Best practice is to use littermate...transgenic strain, the promoter being used should be tested in cell lines to determine how it impacts the expression...chromosomes recombine during meiosis, creating gametes that contain a mix of genetic material from each...recombination at the floxed site, your control should be littermates that express Cre but not loxP. With this two-...

  3. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    Published
    Aug. 9, 2022, 1:15 p.m.
    ...review analyzing and explaining the antibody crisis states that up to 50% of all commercially available antibodies...to more complex brain systems such as non-human primates and humans. Since we need larger-volume antibodies...extensive library of monoclonal antibodies; Addgene collaborates with these companies to create and provide general...includes accuracy, strength, and reproducibility. We test the antibodies on human and mouse tissue and compare... distinct imaged pattern, and exposure times. To test the antibody’s reproducibility, each antibody is...

  4. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ...) to study stem cell differentiation and cancer fates (Singh et al., 2025). Key to this method is their...cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures from days 7, 14, .... (2025). Pre-existing stem cell heterogeneity dictates clonal responses to the acquisition of leukemic...neurons. Created with BioRender.com. Addgene has tested these capsid plasmids in vitro and uses them in...and is endocytosed, the Cre recombinase payload activates a floxed reporter gene (genetically encoded or...

  5. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...surprise and delight,” the very first protein they tested — E. coli expressing good ol’ EGFP — showed a slight...and Maria were skeptical. The team was careful to test the possibility that this might just be an artifact...area measures approximately 82 mm across. Video courtesy of Andrew York and Maria Ingaramo.   MagLOV shows...tools.  Embracing science in the open The York Lab operates with a strong commitment to exploration and collaboration...self-publishing directly on his website (and often posting updates on social media) and freely sharing code, raw data...

  6. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...mechanism of their voltage sensing component, which separates them into two distinct categories detailed below...phosphatases from sea squirts. This design structurally separates the two components of the indicator, which can...was dipicrylamine, a lipophilic compound that distributes itself within the plasma membrane as well. Another...particularly physiologically relevant in neurons, myocytes, and endocrine cells. It follows that the specific...component (i.e., when the dye is lipophilic and integrates into the membrane), the voltage sensing component...

  7. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...it ranks guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At...sophisticated features of CrispyCrunch is how it mutates guide sequences to prevent re-cutting of the sequence... the above are options, the algorithm silently mutates codons in the guide sequence, one by one, from ...

  8. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...quality control, but does not perform functional tests. The newest genome engineer sharing the stage with...be less desirable if one has to target multiple sites in a single experiment. Also, the NGG PAM sequence...nearly 400 times. Researchers have been excitedly testing NgAgo's genome editing applications. So, how does...conducted a survey and asked researchers if they had tested and compared NgAgo head-to-head with CRISPR-Cas9...could not achieve indels with the NgAgo system indicates that it is very tricky to use and fussy to optimize...

  9. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ... plasmid with multiple, individual expression cassettes, having the genes expressed from the same cassette...described below. IRES Elements Translation in eukaryotes usually begins at the 5’ cap so that only a single...originally discovered in poliovirus RNA, where it promotes translation of the viral genome in eukaryotic ... start with a plasmid that has multiple cloning sites flanking the multicistronic element(s), or you could...efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. Kim...

  10. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...biological questions. The two most common uses are: 1) to test the expression level in a specific system (by measuring...occurs when energy transfer from one FP (e.g. CFP) excites the fluorescence of another FP (e.g. YFP). FRET...create the chromophore. This can be from a few minutes after it is translated to a few hours. For example...However, for most FPs it is a few seconds to a few minutes. Photostability can be affected by experimental...greenfluorescentblog.wordpress.com.   More Helpful Websites & Resources: FP guide at Addgene Interactive ...

  11. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...including: how closely homologous the off-target sites are compared to the on-target site, the specific...immediately adjacent to the nearly-homologous target sites. The mere presence of additional PAM sequences should...HR template vector design guidelines? A7: A few notes below are considerations for designing HR donor....with PX330 plasmid, clone in your target guides, test run them, and then select the best guide to co-transfect...site in the donor. For example, choosing target sites that span the knock-in gene. For making mutations...As a successful insertion will destroy the BbsI sites, a double digest should discriminate between positive...

  12. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher HR frequencies...viability of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle...carefully (Lin et al., eLife)! Use a Cas that facilitates HR What if promoting HR was as simple as swapping...It can be! Cas12a is a Cas family member which generates sticky ends upon cutting. Cas12a may promote HDR..., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed repair and termperature-controlled...

  13. Plasmids 101: Colony PCR

    Type
    Blog Post
    Published
    May 12, 2016, 2:30 p.m.
    ...insert-specific sequence. This is a “yes or no” kind of test, with a positive clone amplifying a product and ...primers. These primers are designed to anneal to sites that flank the insert site. A positive clone will...depends on your preferences. Either way, make sure to test your colony PCR primers before using them to screen...Pros Cons Insert-Specific Simple "yes or no" test results Doesn't tell you the orientation of ...of sterile water.  Pick 3-10 colonies in total to test, depending on the number of background colonies ...

  14. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...country.   Figure 1: Number of countries Addgene distributes to per year, 2005-2023. We identified and offered... we contracted with so we can always offer the fastest shipping. However, there were issues that persisted... country are represented in blue; darker blue indicates more views from that country.   While written ... country are represented in blue; darker blue indicates more views from that country. Stats and figures...resources shared via social media, on collaborators’ websites, or as a recommendation in a Reddit thread crowdsourcing...

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

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ...chemiluminescent substrates are available. Labs with limited equipment often opt for the chromogenic substrates since...antibody buffer components can interfere with conjugates. Make sure to do your research before conjugating...sandwich ELISA. In this method, the target antigen competes with a reference antigen for binding to a primary...is non-specific binding or false positives.  If testing a recombinant protein, especially one with tags...

  16. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...factor) promotes editing (Yan, J et al., 2024). They developed a new prime editor that incorporates La’s ...AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation and decoding in pooled...Figure 6: Illustration of split fluorophore assay to test reconstitution via mRNA trans-splicing. A cis-splicing... New and notable Neurodegeneration Collection updates Last but not least, check out the new and noteworthy...

  17. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...typically contains iterons that serve as recognition sites for replication initiation proteins. These iterons...The plasmid does not have many restriction enzyme sites. Hosts often have a restriction barrier as a defense... al. 1977). So if a plasmid has few restriction sites, it may be able to sneak past this checkpoint.  ...incorporate interchangeable antibiotic resistance cassettes (Prior et al. 2010), inducible promoters with ...inducibility (Toh et al. 2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant ...

  18. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...PE6b was very efficient for short, unstructured templates and significantly reduced undesired edits (indels...complex edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing...transcriptional histories, accurate to within 40 minutes. Figure 4: Time-resolved protein ticker ...-PSD-95 [K28/43R] recently passed their initial tests with thick slice (70 μm) immunohistochemistry (IHC...fact, Anti-PSD-95 [K28/43R] has been one of the hottest antibodies in Addgene’s collection since we began...

  19. How to Be an Excellent Trainee

    Type
    Blog Post
    Published
    Sept. 26, 2023, 1:15 p.m.
    ...scientific mentor is typically a professor who operates a research group. Within that research group, ...mean everything! The bare minimum should be taking notes on general protocol steps and other tips and tricks... in the next few days or weeks), look over your notes and protocol(s). Is there enough information there...throughout training, while others prefer to review their notes, process what they’ve just learned, and ask questions...for a copy of the protocol if you prefer to take notes on a printed protocol, or maybe you would like to...

  20. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...initiative and start organizing meetings and take minutes. Get to know your collaborators from other institutions...LinkedIn or ResearchGate allow you to send personal updates as messages and keep you up-to-date on your collaborators...collaborators for example on a holiday or for professional updates. Whenever I see interesting events, articles and...make sure to share them with a short note - this creates a thoughtful way of interacting and is much more..., checking in regularly, and maintaining useful notes that remind me about the current state of a partnership...
Showing: 261 - 280 of 815 results