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We narrowed to 902 results for: Ide

Showing: 541 - 560 of 902 results

  1. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...smaller fragments that are then barcoded for identification. This mixture of barcoded fragments is known...for each sample remain highly normalized across a wide range of input DNA concentrations. This process ...of 96 plasmids on a plate) will typically have a wide range of input quantities. This variation is problematic...are produced from large numbers of samples with a wide range of input concentrations (via proprietary chemistry...alternative to Sanger sequencing because the sample identity information is always displayed at the individual...

  2. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...scarless DNA constructs using homologous regions to guide the joining reaction. BioBrick Assembly will leave...fragments, and generate constructs for producing single-guide RNAs – but it also allows you to skip purification...NEBuilder HiFi so accurate (apart from the high-fidelity polymerase) is the ability to remove, reset and...freely available online. Its simple interface can guide you through both experimental and primer design ...single-stranded synthetic oligo to generate a single-guide RNA (sgRNA). Using only one single-stranded oligo...

  3. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    Published
    Aug. 1, 2017, 1:15 p.m.
    ...Nicholas T., et al. “Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome...sequenced and aligned to the genome to determine genome-wide translational frequency. Although highly useful,...reporter mRNAs. Using translation inhibitors cycloheximide and puromycin, Halstead et al. verified that... translated, the SunTag sites on the nascent polypeptide recruit scFV-GFP, generating green puncta that...

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

    Type
    Blog Post
    Published
    June 1, 2021, 1:15 p.m.
    ...situation. But a few days later we had an idea. And this idea rapidly grew and became Crowdfight COVID-...modern science in our view. And it was this very idea that research is done better with others than by...has been essential to push certain projects. The idea behind the platform was simple: any person working...remote collaborations Read our Transferable Skills Guide article about external collaborations Resources...

  5. Virtual Science Conference Coming Up? Three Addgenies Share Their Experience

    Type
    Blog Post
    Published
    June 30, 2020, 1:15 p.m.
    ...physical location didn’t matter. I didn’t have to consider that aspect and I could plan more with regard ...depending on the conference, the audience can help decide which questions get answered. “You can upvote a...presenting is eye contact. “It’s important to keep your video on and look into the camera to make it seem more...on Twitter where each poster presenter had four “slides” to use for their poster, eliminating the need ...to make space for it,” says Bachle. “On the flip side, you can attend conferences for just a little bit...

  6. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ...efficiencies and expression levels in the cells. Ideally, the chosen reporter is unique to the cell, is ... generations. Stably transfected cells may be considered an entirely new cell line from that of the original..., NIH3T3, CHO, 293HEK, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation 100...hygB HeLa, NIH3T3, CHO, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation. ... 10-14 days depending on how quickly your cells divide. Gentamicin is often used as a supplement in mammalian...

  7. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    Published
    May 27, 2025, 1:15 p.m.
    ...fall back to? We have outlined some important considerations to help you choose the plasmid prep kit that...or by incorporating recycled paper and plastic. Aside from the more obvious swaps like using smaller bottles...Where can you look for help?  All manufacturers provide specs, protocols, and quality control information...help explain the rationale behind each step and provide troubleshooting advice. In these cases, it can ...between these manufacturers, as price is a large consideration for most labs. Many institutions have preferred...department for these.  While budget is an important consideration, you do want to make sure you’re getting the...

  8. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    Published
    April 27, 2023, 1:15 p.m.
    ...Visit Our AAV Guide Page Find Ready-to-use Virus for Your Research Visit Our Lentivirus Guide Page ...understand brain function So far scientists have identified 180 brain regions. Both this geography and the...Using retrograde AAV AAVrg can be used to deliver a wide array of genetic tools for neural interrogation ...where the cell bodies of the projection neurons reside.   Impact for scientists While the mechanisms ...

  9. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    Published
    July 22, 2025, 1:15 p.m.
    ...chemical substrate (and sometimes cofactors) to be provided and are generally dimmer than fluorescent proteins... remains popular, other luciferases have been identified in the sea pansy (Renilla reniformis), click ...assays (low background, no photobleaching) but provides flexibility by pairing the luciferase with a variety...change in conformation of a single protein. Considerations for luciferase assays Luciferases can only ...luciferase output at different wavelengths. Addgene provides empty backbones with the luciferase gene, a variety...

  10. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...of dNTP in the reaction mix, the researcher can decide exactly how far the T4 exonuclease proceeds: at...at the first occurrence of the chosen nucleotide, T4 will switch to polymerizing and then stall due to...the “damage” and producing a complete plasmid. Besides being compatible with any vector, the SLIC protocol...multi-fragment DNA assembly, and its low cost makes it ideal for researchers doing large amounts of cloning. ... 10.1007/978-1-61779-564-0_5. PubMed. JBEI-J5’s guide to SLIC Resources at Addgene Ligation-Independent...

  11. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...in the lab. This is commonly done using calcium chloride which permeabilizes the cell membrane so the bacteria...inject their genetic material into the cell. Once inside, phages can follow one of two different life cycles...both of these life cycles bacterial DNA can be accidentally packaged into the newly created phages. Transfer...referred to as transduction. Transferred DNA once inside the infected bacterium can either exist as transient...Addgene’s protocol page, which has both protocols and videos for techniques in basic molecular biology, plasmid...

  12. Antibodies 101: Flow Cytometry Controls

    Type
    Blog Post
    Published
    June 6, 2024, 1:50 p.m.
    ...particular marker, you can use the reference plots provided by the antibody companies or check the literature...sample. The isotype antibody will be virtually identical to the antibody for your marker of interest. It...for that particular marker. This enables you to identify any non-specific binding to know that your positive...unsure what amount of antibody to use, you should consider using an isotype control. Pro tip! Many companies...Cytometry Resources on Addgene.org Addgene's Antibody Guide Browse Addgene's Antibody Collection        ...

  13. Communicating Your Science With Help From ComSciCon

    Type
    Blog Post
    Published
    July 19, 2018, 1:12 p.m.
    ...suggests a change to a slide, think about whether the same principle applies to other slides in your deck.  If... craft sentences for abstracts and presentation slides, learn from audiences as they do science outreach...of a new science communication initiative that provides yet more scientists with opportunities to practice...Resources on Addgene.org Check out how Addgene uses videos for science communication ...

  14. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    Published
    Nov. 19, 2019, 2:08 p.m.
    ...more OptoNBs would allow for more uses both inside and outside cells. References Gil, Agnieszka A., et...exciting because using light to control signaling provides greater spatiotemporal resolution than pharmacological...domain of the Sons of Sevenless (SOS), a guanine nucleotide exchange factor that activates Ras. They expressed...more about optogenetics in Addgene's optogenetics guide ...

  15. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    Published
    June 29, 2021, 1:15 p.m.
    ...protein. Animals are first injected with a small peptide that represents a short, unique amino acid sequence...isolate individual hybridomas and ELISA is used to identify the hybridomas producing the most specific antibodies...antibody, meaning that the antibodies are largely identical between lots.  Disadvantages of monoclonal antibodies...antibody-encoding genes over time as the hybridoma cells divide. These changes can alter the antibody that is actually...produced by the hybridoma. Any genetic drive can be identified by sequencing the heavy and light chains. To ...

  16. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    .... To make a point mutation, you design two oligonucleotide primers with the desired mutation, then use...3-4 phage proteins are needed and these can be provided via a separate plasmid or be inserted into the...insertions and deletions. The figure above shows guidelines for primer design; a desired insertion should...REPLACR-mutagenesis successfully incorporated a 60-nucleotide insertion; beyond this insertion size, primer...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which are...

  17. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    Published
    July 28, 2015, 2:30 p.m.
    ... in the human genome. The pair has focused considerable attention on transcription factors, starting...have good antibodies. The results look almost identical. Addgene: What advantages does your system have... also easy to run things in parallel. Addgene: Besides being fast and efficient, what other challenges... Myers: Only a small percentage of the proteins inside a cell have a commercial antibody against them.... to us to get them. Even though we’re happy to provide them, that never works on a large scale. The world...

  18. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...blogger Didem Goz Ayturk with edits and updates from Addgenies Karen Guerin and Susanna Stroik. Didem Goz ...administered in an organism. These features make AAV ideal for human therapies and in vivo laboratory experiments...therefore the transduced gene is episomal and does not divide and replicate in the same way the host DNA does...Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All Viral Vector Resources...

  19. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...thousands of plasmids, using both mass production genome wide screens and the more traditional “one at a time”...time. We also handle the MTA, QC the plasmids, provide helpful resources, and, on the rare occasion things...organized lab saves time and money across the board. Consider alternative cloning strategies There are many ...either don’t scale or have limited scaling. The downside of high-throughput work is that you give up a ...engagement with and use of repositories like Addgene provides large-scale savings. How large? Using Doench’s...

  20. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...affects protein function (Bornhorst 2000).  One downside of polyhistidine tag-based purification, however...contamination, such as mammalian production systems, consider using these more selective substrates along with... containing a polyhistidine tag, you’ll need to decide whether to add the tag to the N- or C-terminus ...the opposite end.  If neither placement works, consider purifying under denaturing conditions. Unlike ...References  Resources on addgene.org Addgene Antibody Guide Ready-to-use antibodies from Addgene More Resources...
Showing: 541 - 560 of 902 results