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  1. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    Published
    Feb. 2, 2018, 3:20 p.m.
    ...things or communications planning.” To beef up her resume, she decided to complete a certificate in Strategic...Podcast Episode on Science Communication Check out Other Science Communication Posts Resources on Addgene.org...In this post of the Careers in Science Communication blog series, you’ll hear from Susan Keown, a staff...Cancer Research Center. Susan Keown’s science communication journey started with something unexpected: termites... time position with a Washington D.C.-based communications consulting firm, Palladian Partners. “I was...Strategic Communications and Public Relations at the University of Washington and the connections she made...storytelling is an impactful tool for science communicators. “The power of a story is to make a human connection...

  2. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ... to help decide whether work is suitable for publication in their journal. The early peer-review attempts...information that would prevent the study to be replicated or omission from the literature. I think that...Addgene.org Learn about depositing plasmids pre-publication Find molecular biology protocols Find eBooks ...

  3. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    Published
    July 26, 2018, 12:46 p.m.
    ...phosphatase-accessible. Kinase Translocation Reporters (KTRs) Kinase Translocation Reporters (KTRs) are genetically...provides an alternative tool with their Kinase Translocation Reporters (KTRs) whose cellular localization...activity is enhanced, leading to cytoplasmic translocation of the reporter. As a result, the KTR is shuttled...nuclear export and import rates can influence KTR translocation. Work from the Covert lab suggests that these... negligible effect on KTRs, however nuclear translocation rates are another variable to consider when ...measurements of kinase activity in single cells using translocation reporters. Nature protocols, 13 1, 155-169. ...

  4. Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

    Type
    Blog Post
    Published
    April 12, 2018, 1:02 p.m.
    ... Our education partnerships have grown to include local school districts, professional education associations...contributed by guest blogger Courtney Price, the Education & Outreach Specialist for the Arabidopsis Biological...resources for Arabidopsis and related species. Located at The Ohio State University in Columbus, Ohio,... more than 100,000 samples to researchers and educators in 60 countries each year. Introducing Arabidopsis...outreach program kicked off with the release of 20 education kits designed for use in K-12 and college-level...Translating Research on Arabidopsis Into a Network of Educational Resources). Arabidopsis is used in research to...blogger, Courtney Price!  Courtney Price is the Education & Outreach Specialist for the Arabidopsis Biological...

  5. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ...voltage indicator Archon Article contributed by Shreya Vedantam  Listen to the voltage indicator podcast... genetically encoded voltage indicators. To expand the voltage indicator toolbox, Ed Boyden’s lab developed...customized for protein visualization, relocation, degradation, modification, and scaffolding. In their study...xCas9, and requires no additional chemicals or modifications. This versatility opens up the potential for...Archon1, a novel opsin-based fluorescent voltage indicator. Archon1 can embed itself into the cell membrane...response to blue light (445nm). The discovery and applications of protein partners that bind to only the light...

  6. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ... writing for the Addgene blog, sharing Chemistry Cat memes was how I used social media as a scientist....needed to start using Twitter. Sharing Chemistry Cat memes use to be how I used social media as a scientist...remotely as an Addgene Science Communication Intern. Developing science communication skills, while also learning... on a panel about “Social Media for Science Communication” at the American Society for Cell Biology (ASCB...Twitter Learn about using Reddit for science communication ...

  7. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...GpNLuc and OgNLuc are available through Addgene (Cat.# 70185 and 70186, respectively). Antares Chu, J...construct for Antares is available through Addgene (Cat. # 74279). Enhanced Nano-Lanterns  Suzuki, K., et...et al (2). In this post, I’ll cover two great applications of the NLuc technology. The first couples NLuc...fluorescence. The paper reports a wide variety of applications of the Enhanced Nano-Lanterns including monitoring...2017) Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging...

  8. Cultivating Community Science at BosLab

    Type
    Blog Post
    Published
    Nov. 2, 2017, 1:35 p.m.
    ...Experimenting in an Open Source Lab: from CRISPR to Cats Stem Cell Models for Disease & Open Science: Interview...state of Massachusetts. Because we’re conveniently located next to a major Biotech hub, building up our lab...pipetting experience. However, after a few months of dedicated time at BosLab, he became our bacterial transformation...projects range from hacking probiotics to filming educational lab videos, while some are looking to develop...microbial diversity in different soil conditions/locations but to engage the local community (including local...Order Plasmids from Addgene! Find Free and Open Educational Resources at Addgene! ...

  9. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    Published
    Dec. 20, 2018, 1:15 p.m.
    ...) in an old Girl Scout Cookie factory with their cats, Harley and Lexi, and a clutter ofoutdoors-only ...with healthy mitochondria, with the intent to eradicate mitochondrial diseases. The key success of the...

  10. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...found climbing mountains or playing with his two cats. Keira Havens grew up in Hawaii where she was fascinated...biotechnologists face is public perception of genetic modification (GM) and synthetic biology. The mistrust and...cannot build what we do not understand, and the complicated and unique metabolic processes of plants are ...

  11. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    Published
    June 5, 2014, 3:32 p.m.
    ...found climbing mountains or playing with his two cats. Keira Havens grew up in Hawaii where she was fascinated...plants engineered to detect explosives and rapidly indicate their presence. They are genetically modified ...enjoy and see a future in. And we decided to make education and outreach a cornerstone of our business, because...

  12. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...spending time with her horse, and wondering what her cat is thinking. Find her on Twitter @JessicaSacher. ... whole genome amplification (by a mechanism known as multiple displacement amplification) even in the ...same location that their host colonies grew. If your cells of interest stick to the same location as a...adding methyl groups, glycosyl groups, and other modifications to their bases in order to protect their DNA...DNA from host cell nucleases (4). To further complicate matters, even when sequenced, phage genomes tend...compressed nitrogen gas; this shears DNA at random locations and allows for a given fragment size to be selected... for physical shearing exist as well, such as sonication and needle shearing. Essentially, any method ...

  13. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ...Origin of Replication? See our Plasmids 101: Origin of Replication blog).  The replication problem Plasmids...directly inhibit replication by binding to the origin of replication and blocking replication machinery (del... of microbial plasmids: classification and identification based on replication and transfer systems and...plasmid that contains the origin of replication and the replication control machinery (Need a refresher... that inhibits replication when copy number in the cell is high, but allows replication when copy number...either indirectly inhibit replication through inhibiting translation of replication machinery proteins, or...antisense RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated...

  14. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    Published
    June 9, 2022, 1:15 p.m.
    ... can and do consume science communication.  Formal education is education that is provided through a structured...underlying all communications efforts.  Scicomm definitions (as used by Addgene)    Science communication, or science...time engaging with science communication efforts.  Scientific communication or scientific writing is peer...to note that it is the actual communication itself, not the communication method, that determines if something...scientific or science communication. A Tweet is still considered scientific communication if the language is... engage or disengage at will.  Informal education is education that is provided, typically ad hoc, for...Series, where we’ll be talking about science communication, best practices in the field, and even getting...

  15. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...epigenome (the sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting...DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic... the catalytic domain of p300 acetyltransferase increases levels of H3K27ac histone modification at specified...Preps for Epigenetic Modification Why use epigenetic modifiers? Epigenetic modification is certainly not ...Nature Communications 8: . https://doi.org/10.1038/ncomms16026 Find plasmids from this publication at Addgene..., 2017 and updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...

  16. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    Published
    Dec. 12, 2019, 2:22 p.m.
    ...that allows category-specific, narrow searches. We defined three top level categories — catalog, educational... in a given category, and serve as a shortcut to a search scoped to that category. Catalog ID suggestions...prefer to browse and discover items in our catalog or educational resources through our main dropdown menu... of Addgene’s customer’s needs. Categories We created a categorization of our information in order to ...depositors — each with subcategories such as plasmids and viral preps for catalog, blog posts and protocols...80,000+ reagents and 700+ educational resources — including blog posts, protocols, guides, and collections...sequencing data; as a result, the materials in our catalog are very well-characterized. During this time, ...

  17. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    Published
    Feb. 11, 2014, 12:13 p.m.
    ...the grant application process. When my PhD advisor first asked me to write a grant application due the ...in a previously funded grant application to see how funds were allocated. Step 3: Get feedback from your...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...advisor alone applies for these competitive grant applications; however, in some cases, your advisor may choose...writing, volunteered to take the lead on various applications throughout my graduate career, and helped receive...Proposal writing: Improves technical writing and communication skills, Elucidates the big picture and future...

  18. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...or DYKDDDK 1 Affinity and Purification Good for antibody-based purification; has inherent enterokinase...site GST Large Protein 26 Purification and Stability Good for purification with glutathione; protects...used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity and Purification Very small...GKPIPNPLLGLDST   1.4 Affinity and Purification Good for antibody-based purification Combo and cleavage tags...multitude of uses including (but not limited to) purification, detection, solubilization, localization, or...fusion proteins to aid in the expression and purification of recombinant proteins. Tags for stability...be fused to recombinant proteins for one-step purification with glutathione, can also protect against proteolysis...

  19. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...single peak, indicating one amplicon and no primer dimers. (b) A shoulder peak indicating primer dimers...Method Standard Curve?  Amplification Efficacy Calculation Amplification Efficacy Assumptions ...understanding and technical proficiency with common applications - now let’s dive right in! You know qPCR is...is most commonly used to measure relative quantification of mRNA, which can be used as a proxy measure...as it solves, so understanding and using the application correctly is imperative. Picking your chemistry... at least 25 bp away from each primer, ideally located close to either the forward or the reverse primer...dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K. Filutowska; Selma...

  20. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...variety of projects in the space of science education, communication, and policy. In this post, I hope to tell...science policy and science communication A career in science communication or policy requires broad interest...pursuing a career in science communication or policy, being able to communicate your work to a non-technical...series on careers in science communication Find all of our science communication posts Extracurricular activities... of New Initiatives for the Personal Genetics Education Project (pgEd). Advances in genetic technologies...inequalities and injustices. The Personal Genetics Education Project (pgEd), where I currently work, strives... as well as the personal, social and ethical implications of genetics. In these conversations, pgEd strives...

  21. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...switches. Stabilizing gRNA modifications  Sugar phosphate backbone modifications The recognition sequence...phosphorothioates with 2’-O-methyl modifications, proved more stability than just one modification alone (Hendel, et al...ordered as synthesis modifications when purchasing your gRNA. Pro tip: modifications are typically incorporated...stabilizing modifications listed above were developed before CRISPR-Cas9, for use in applications like siRNA...worrying about the status of your RNA. gRNAs modifications are even going beyond stability, adding color...cellular RNA molecules with post translational modifications. One of the most common of these is 2’-O-methylation...nucleolytic attack. Another stabilizing sugar modification, found at the same site, is 2’-fluoro (2’-F)...

  22. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...FAIRE-seq, and CHIP-seq), genome-wide identification and quantification of enhancer activity have remained...compartments (cytosol, Addgene catalog #71245; mitochondria, Addgene catalog #71247) Rosella provides an ...CpG methylation by fusing catalytically inactive Cas9 (dCas9) with the catalytic domain from DNA methyltransferase...  Superglue proteins with SpyTag/SpyCatcher and SnoopTag/SnoopCatcher The development of recombinant DNA...alternating the use of the SnoopTag/SnoopCatcher and SpyTag/SpyCatcher pairs. In order to improve the ease...MBPx-SpyCatcher construct. This construct is used to produce a maltose binding protein/SpyCatcher fusion...of larger fusions through SpyTag/SpyCatcher and SnoopTag/SnoopCatcher linkages. Once all of the desired...

  23. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...higher education nonprofit entities related to or affiliated with institutions of higher education nonprofit...for self-sponsorship of the I-140 application meaning that the category does not require a foreign national.... NIW requirements stress “educational background, expertise, publications, and plan for future activities...received 116,224 employment based green card applications and approved 91,023 during the first 3 quarters...for a role that is closely aligned with their education/experience, i.e. a research role, for a Ph.D. ...well-tailored to meet the needs of researchers. The categories are as follows: Cap Subject Industry H1B Academic...occupation" as requiring theoretical and practical application of a body of highly specialized knowledge in ...

  24. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...in PubMed and choose the appropriate publication. This publication will be listed at the top of the final... with publication details or permission to distribute, or Addgene finds your journal publication describing...classified in multiple categories, please check all of the appropriate categories (e.g., for a lentiviral... "Published plasmids" or "Unpublished or pre-publication plasmids" and set a desired distribution status...quickly associate your plasmids with the original publication where you described them. Search for your article...plasmids. You can submit "Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that...accepted. Addgene also encourages submitting pre-publication plasmids so they can be available when the paper...

  25. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...affinity purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes...genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as dCas9 (Jinek ...interest targeted by a gRNA. CRISPR-mediated purification of a specific genomic region In 2013, the Hodaka...Subsequent mass spectroscopy studies allowed for the identification of proteins associated with the IRF-1 locus...steps (Fujita et al., 2013). CRISPR-mediated purification of a specific genomic region alleviates these...the initial report describing CRISPR-mediated purification of a specific genomic region, numerous adaptations...different fields of study. In addition to the identification of DNA binding proteins, this method can characterize...

  26. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...Technical Tips). Minigenome replication and transcription In order to replicate the minigenome, the viral...transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems...allowing researchers to study aspects of the EBOV replication cycle under BSL-2 conditions (4). What is a ...that are crucial cis-acting signals for genome replication and viral gene transcription (6). The EBOV genome...5’ ends of the genome which are required for replication and transcription (Figure 1). All viral genes...cis-acting signals necessary to direct minigenome replication and reporter gene transcription by the EBOV polymerase... in 2, 5). The minigenome is capable of being replicated and transcribed in the cell when the appropriate...

  27. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    .... Tracking samples - it's all about location, location, location There are many different types of samples...but we also have a dedicated and detail-oriented lab team that works carefully to catch mistakes. Each sample...assigning a physical location to your samples, include as much detail about that location as possible. Are...sample a unique location in a specific freezer will help ensure that samples are easy to locate in freezers...also help us keep track of the tubes’ physical location in the lab. These barcodes additionally allow ...Addgene. Each sample at Addgene has four layers of location which can all be extrapolated from the 2D barcode...Plate Freezer Rack Freezer Tubes are assigned a location within a plate that can hold up to 96 tubes (see...

  28. 6 Tips for Grant Writing

    Type
    Blog Post
    Published
    Oct. 18, 2016, 2:30 p.m.
    ...Series 4. Become a science communicator More and more fellowship applications are asking for a 200-300 ...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...someone else in the department) to read over your application and give you feedback. They’ve most likely been...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be ...

  29. Antibodies 101: Affinity Tags

    Type
    Blog Post
    Published
    Sept. 19, 2023, 1 p.m.
    ...cleaved after purification, so as not to perturb protein function for downstream applications. Tag cleavage...thoughtful planning, affinity tags can make protein purification a cinch. Types of affinity tags, the proteins... Affinity tags are used primarily as protein purification tools (and can have secondary functions as detection...single protein is expressed, with the intent of purification, it will be expressed along with all other cellular...available antibodies, resins, beads, and other purification aids readily available. Choosing the best tag...tag to remain on your protein for downstream applications Below, we describe a few of the most common...cleavage must be performed after primary purification and requires a second clean up step to remove the cleaved...

  30. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...serially transfected using a cationic vehicle. The following synthetic modifications are used to protect the... to verify there’s no genomic integration Replicating EBNA1 episome Non-integrating Medium Medium ...creates heterogeneous iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing...Retroviruses are not considered safe for clinical applications.     Lentiviral: Lentiviral vectors infect dividing...lentiviral vectors insert in different genetic locations in each transduced cell, so there’s unavoidable...unavoidable heterogeneity between clones. This can complicate comparisons made between samples, i.e. transgene...extent of reprogramming could be affected by the location of insertion. Lentiviral insertion can also disrupt...

  31. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various hosts...the ori and allow for replication. However, if plasmids have their own replication machinery such as Rep...different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow... while others have a broad host range and can replicate and stably maintain the genes they carry among.... These features are: The plasmid origin of replication (ori) contains structural elements that are versatile...structure allows plasmids to utilize diverse host replication machinery. For instance, the ori typically contains... iterons that serve as recognition sites for replication initiation proteins. These iterons vary in their...

  32. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ...their CV or grant applications. A number of organizations (such as ASAPbio) advocate for preprints, and...the most important part of your research is communicating the results of the work. Yes, those control ...any impact! There are lots of ways scientists communicate with each other: from formal mechanisms like ...poster or talk, or submitting patent or grant applications; to informal settings like lab meetings, social...of the most significant shifts in scientific communication is the rising use of preprints, or scientific...publishing in a journal.  Accelerating scientific communication Preprints have been common in fields like math...under a CC BY-SA license. Many scientists and advocates have argued that while peer review is valuable...

  33. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...desired modification. Finally, the positive clones are sequenced to confirm the desired modification and ...polymerase activity (for amplification), 3’->5’ exonuclease activity (increases amplification fidelity), and no... genome modifications. Occasionally, multimerization of the PCR primers can cause duplication of the primer...the desired modification have been identified (and confirmed to be free of primer duplications), final validation...and the absense of additional modifications.   Site-directed mutagenesis of Plasmids. A) Generation..., which flank the targeted region. Upon PCR amplification, the template is removed by DpnI restriction...extension time to match the size of your template. Amplification of GC-rich plasmids is facilitated by the addition...

  34. Summer SciComm Series: Master of SciComm

    Type
    Blog Post
    Published
    July 21, 2022, 1:15 p.m.
    ...begun my journey towards a career in science communication, an established field of inquiry with which ...University of Otago’s Department of Science Communication the following year and, having thoroughly enjoyed...straight on to a master's degree in science communication. The first year of the course involved a whirlwind...world of industry. Along this road, science communication graduates are often well-equipped. Many institutions...truly benefit others. Herein, a good science communicator—with an understanding of how to appeal to a ...pursuit of public engagement. Studying science communication is an equally fruitful endeavor for those embarking...towards academia. Many of those entering science communication (myself included) come from a scientific background...

  35. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    Published
    Aug. 18, 2023, 4:15 p.m.
    ...methods of replication. In fact, an entire classification scheme, known as the Baltimore replication classes...that in a bit! Viral genomes and replication When it comes to replication, viruses tend to march to the ...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit: ...an intermediary DNA replication step between DNA and mRNA. DNA genome replication can involve an RNA or...cell. The cell then uses its own machinery to replicate the viral genome and proteins. This is, for the...before being translated into proteins. RNA viruses replicate their genomes using a variety of RNA or DNA intermediaries... bacteria and eukaryotes, which use a binary replication process whereby one cell copies itself and “splits...

  36. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...would be a good time to talk about the amplification and verification processes we use to ensure high-quality...your own experiments. Library amplification 101 Library amplification seems straightforward: E. coli ...transformed with the library DNA, they grow and replicate, and then their DNA is harvested, much like one...library has its own depositor transformation and amplification protocol, but the overall workflow is similar...ll need to remember when undertaking library amplification and sequencing. Maintain representation of the...electrocompetent Stbl4 cells when performing amplifications, and we use electroporation to ensure a good...your sequencing results represent a successful amplification? To provide a point of reference, Addgene commonly...

  37. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...potential applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves... the perfect voltage indicator and optogenetic actuator pair. Ideally, indicators should possess fast ...random genomic location than to express the tagged protein from a defined genomic location or from its endogenous...temporal control over CRISPR/Cas9 mediated genome modification and gene expression. The plasmids associated... system widely accessible for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol ...other ions In BRET, A mutant Renilla luciferase, catalyzes the oxidation of its substrate, luciferin, which...amenable to both high and low throughput screening applications. The system can be readily modified to use different...

  38. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...them. An easy way to do this is gel purification. In gel purification, you use a voltage difference across...backbone. Sample results indicative of successful and unsuccessful ligations are indicated below. A successful...grow overnight cultures for DNA purification. The simplest purification you can do is a miniprep, but if...digests. Of course there’s much more detail and verification required for the process to work well, so let...backbone. Because you lose some DNA during the gel purification step, it is important to digest plenty of starting...prior to the ligation step or prior to the gel purification step, depending on the phosphatase you choose...   2. Isolate Your Insert and Vector by Gel Purification     Now that you’ve cut your insert and vector...

  39. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    Published
    July 13, 2023, 1:15 p.m.
    ...you.  Starting from plasmids requires amplification, verification, and lentiviral production steps before...the same proportion after amplification as they were prior to amplification. And as anyone who has worked...process for using pooled lentiviral libraries. A) Amplification, validation, and lentiviral generation of a ...pooled library is transformed into E. coli for amplification. Transformed E. coli are allowed to grow on ...into HEK cells allows production of lentivirus. Purification of this lentivirus results in the pooled library...//pubmed.ncbi.nlm.nih.gov/27366891/   Amplification When you request a DNA pooled library from Addgene...might receive a similarly-sized aliquot from an amplification done by Addgene.) This small aliquot is not ...

  40. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ...are used across species: replication, selection, and expression. Replication It’s usually not enough to...chromosome has a single origin of replication (ori), where DNA replication begins, so a bacterial plasmid...Eukaryotes are a bit more complicated. Yeast vectors may contain an autonomously replicating sequence (ARS), a...fast-growing populations of cells, the vector needs to replicate alongside its host, or it will be lost from the...compatible ori that will be recognized by its host’s replication machinery. A shuttle vector designed for unrelated...experiment. Yeast shuttle vectors have these replication sequences for their yeast targets as well as ...episomal vectors is to include viral oris to drive amplification, which helps increase cellular uptake, but this...
Showing: 41 - 80 of 833 results