Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 273 results for: plat

Showing: 41 - 60 of 273 results

  1. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...antibiotic resistance marker. After transformation, plate the cells on medium containing the corresponding...contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing agar. Only cells... are auxotrophic for specific amino acids. When plated on media lacking the essential amino acid, the ... a toxic metabolite in gram-negative bacteria. Plating on sucrose medium will select for cells that contain...gene is used for negative selection on 5% sucrose plates. Exchange of the sacB gene for the gene of interest...

  2. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...DNA from each well of a 96-well plate. Then, samples from several plates are pooled together to create ...samples individually, plexWell™ reduces every 96 well plate to a single tube after the first tagging step. As... samples (e.g. a collection of 96 plasmids on a plate) will typically have a wide range of input quantities... from plexWell™ NGS data. Once Addgene produces plates of plasmid DNA, they are shipped to seqWell, where...

  3. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    Published
    Feb. 28, 2023, 2:15 p.m.
    ...devices for research. There are a number of new platforms (such as openUC2, which I am a co-founder of),...that make everyday microscopy much easier. On platforms such as Github, users can download the software...example, can also be printed quickly in Madagascar. Platforms that allow for on-going creativity with the construction...organizations and companies to provide problem-solving platforms.     Fig. 3: Different workflows involved...Prakash, "Squid: Simplifying Quantitative Imaging Platform Development and Deployment." bioRxiv. 2020. Diederich...

  4. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ...engineers regularly use the platform, why scientists are using the platform, and tips for getting the most...from Nature Jobs Blog While Twitter is a popular platform for scientists, it’s not the only tool in a scientist... Facebook. For each of the three social media platforms, there’s some statistics on what percent of scientists...given you tips for using various social media platforms, and 3) helped you fit social media into your ...

  5. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...reverse transcriptase template, starting with about 10–16 nucleotides. For longer templates, it is even more...to the genome and a reverse transcriptase (RT) template including the desired edit. Figure 1: Prime...primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com. The first...encourage the cell to use the edited strand as a template during DNA repair. Test multiple nick sites, starting...

  6. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...secondary nanobodies blog post.  A rapid selection platform for generating sybodies against membrane proteins...weeks (Zimmermann et al., 2020). This selection platform was designed so that any standard lab can quickly...in the form of mRNA for academic research. The template plasmids for generating your own libraries are... out with anti-Myc antibodies bound to an ELISA plate to capture Myc-tagged sybodies. Then, the biotinylated...proteins. The full protocol for this sybody selection platform can be found here and relevant plasmids are available...

  7. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...shaking at 37 degrees) can be shortened since the plated cells will have time to produce beta-lactamase ...to produce satellite colonies when used on agar plates due to the fact that beta-lactamase is excreted...Cons: Prone to satellite colony formation on agar plates, less stable at high temperatures and in acidic... and is less susceptible to degradation in agar plates. This prevents satellite colonies from forming ...Molecular Biology Protocols Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog...

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

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...then recognizes the encapsidated minigenome as a template for replication and transcription, as it would...-minigenome). The anti-minigenome is used as a template by the viral polymerase to synthesize additional...addition, the polymerase will use the minigenome as a template to transcribe the reporter gene (e.g. firefly ...polymerase complex and utilize the minigenome as a template. Thus, it is important to choose cell lines that...and 2 μg minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...

  9. What's New in CRISPR - May 2019

    Type
    Blog Post
    Published
    May 28, 2019, 12:49 p.m.
    ...genome-wide CRISPR inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9 CRISPR interference...inhibition library Covalent tethering of DNA repair template to Cas9 Homology-directed repair (HDR) can be ...using a single-stranded oligodeoxynucleotide HDR template covalently attached to the Cas9-gRNA RNP complex...Candida albicans A new CRISPR interference (CRIPSRi) platform for targeted gene repression in Candida albicans...

  10. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...establish a next-generation sequencing (NGS) QC platform that allows us to provide scientists with complete...launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral vector (...ligation and tagmentation require a double-stranded template.  Using viral genome sequencing to verify the ...double-stranded species that could be used as a template for downstream applications (Berns, 1972). Using...

  12. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...without double-strand breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime editing...increase activity, enhance binding between the template and primer binding site, increase processivity...remake that strand using the edited strand as the template. The PE3 system does just this by including an...PE = prime editor; RTT = reverse transcriptase template; PBS = primer binding site. Image from Doman et...extended 3′ tail containing the reverse transcriptase template and primer binding site. If this tail is degraded...

  13. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    Published
    Feb. 8, 2022, 2:15 p.m.
    ... to glance at as you work. Draw your plate maps onto your plates, alongside your physical copy. If you...reminders on a sticky note   Visual Planning  Plate maps, pre-labeling tubes, and even drawing out experimental...example, setting up your pipette tips to match your plate layout can help you track samples as you add them...

  14. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...desired mutation, then use those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested...nick repair. This method can work well, but poor template digestion can result in a high number of background...mutations, to amplify both strands of a plasmid template. In addition to containing the mutations, the ...After being treated with DpnI to digest away the template plasmid, these PCR products are directly transformed...

  15. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...samples through our lab using barcoded tubes and plates. The critical pieces of information for a given...100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then ... database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for forward...shipped along with their corresponding glycerol plate on dry ice to the sequencing provider. Each sequencing...

  16. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...genes. For this you would need to provide a DNA template to repair the double strand break via homology...Cas9/sgRNA and one vector for your specific DNA template. Using this two vectors system, Manuel Gonçalves...This technology therefore represents a potential platform that could be realized via simple switching of...tools can also be used to deliver Cas9 and DNA templates in somatic cells of adult animals in order to ...

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

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

  18. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ... need to deliver the Cas protein altogether. Platforming AAV Interest in interrogating and mapping the...glioblastoma, and generate programmable screening platforms for mouse tissues. Your AAVancement could be next...highly scalable cell type-specificin vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https..., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. J. (2023). Transcriptional linkage analysis ...

  19. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...homologous recombination using a repair template. These templates can include loxP sites to create conditional...recombination. The concept is simple: deliver a template with a modified version of the targeted sequence...sequence to the cell which will recombine the template with the endogenous gene. Here, we'll describe the ...

  20. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ... from an LB plate. In my hands, a 5 mL starter culture inoculated from a colony on a plate will reach ..., check out Harrison et al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase...difficult. Don’t leave your plants on selective plates for too long: even resistant plants will be affected...
Showing: 41 - 60 of 273 results