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Showing: 41 - 60 of 415 results

  1. Increasing Your Work's Visibility with Addgene: Citation, Search, and Collections

    Type
    Blog Post
    ...request that you cite it using both its Addgene ID number as well as the reference for its original publication...plasmid from Addgene using its unique Addgene ID number, your publication will be listed on our website...-  While it is tempting to rely on the citation number when comparing two plasmids, we recommend reviewing...can search for plasmids by name, PI, Addgene ID number, or keyword. Although an initial search might give...in the repository, our scientists have curated a number of special collections pages. These pages serve...

  2. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...spinning disk microscopy, a metal disk containing a number of holes rotates through the excitation light path...axial volume of a sample. Therefore, when a large number of optical slices are acquired all fluorophores...in the final image. Over the past two decades a number of techniques have been developed that allow for...example is the lattice lightsheet that projects a number of thin cone-shaped beams into the sample. These...require optical sectioning, but open the door to a number of other techniques that can improve contrast and...when imaged. Super-resolution microscopy uses a number of optical and chemical ‘tricks’ to turn subsets...

  3. Addgene Depositors Get More Citations

    Type
    Blog Post
    ...Journal Weighting Example Article Number (A) Year Published (B) Number of Citations (C) Journal Name (D...its vast network of articles to track the total number of citations the paper receives. From the database...correlation, but it wouldn’t be causal. To control for a number of differences between those papers with Addgene... see the full impact from using these weighted numbers. Note: this does drop some journals from our dataset...

  4. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...our partnership with Addgene to sequence large numbers of plasmids. plexWell™ technology overview For...pooled library prep”, in which a relatively large number of samples are subjected to a DNA-barcoding step... problem often arises from the fact that large numbers of individual input DNA samples (e.g. a collection...time and consumables, can be daunting for large numbers of samples. As mentioned above, one of the primary...that normalized libraries are produced from large numbers of samples with a wide range of input concentrations...

  5. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...regulation of bacterial ORIs determines plasmid copy number within the bacterial cell. Similarly, the specific...yeast cell. These elements control not only the number of plasmids found in each cell, but also whether...+ copies to stably propagate per cell. The copy number of these vectors can also be controlled if specific...than normal physiological levels due to high copy numbers. This creates a potential metabolic burden on the...thus allowing the yeast to maintain higher copy numbers. Additionally, improvements in antibiotic selection...

  6. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ... a specific number of fragments, you can also ask it to give the maximum or minimum number of fragments...complete with predicted fidelity. You can specify number of overhangs, three or four base-pair sequences... it into multiple fragments. You can define the number of fragments, and the “search windows” you’d like...types, circular assemblies yielded 500 times the number of plaques than the linear ones. The 52-piece circular...

  7. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...properly, it is unstable and may only work for a small number of qPCR runs before a new one is needed. For these...of the target DNA. This partitioning reduces the number of PCR inhibitors per reaction and allows for enhanced...control wells. You’ll also notice a reduction in the number of positive droplets as the dilutions increase....here are a few things you can try: Increase the number of cycles. After additional rounds of amplification...PCR is well suited for the detection of low copy numbers. For this reason, it can be used in the detection...

  8. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...able to include specific plasmid ID numbers. If you know the ID number for a plasmid, your straightforward...own CSO. But what if you don’t have plasmid ID numbers, or you aren’t sure if any plasmids from a paper...potential results. Just as in the previous layout, the number of plasmids matching each filter is displayed in... Only see plasmids with more than the indicated number of requests Availability to Industry: Restrict ...

  9. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...fusion.  The former allows for screening greater numbers, but the latter eliminates the step of identifying...leading to detectable reporter signals depend on a number of factors, including protein expression and correct... or more of these parameters can lead to a high number of false negatives in Y2H screens. Testing your... a wide range of prey proteins, there are now a number of two-hybrid systems available in model organisms...and produce a more vigourous screen is to use a number of different bait and prey vectors. This has been...

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

    Type
    Blog Post
    ...efficacy in any given system can be affected by any number of factors, including the age of the model organism...and targeting The amount of vectors available and number of requests tell only part of the story. Viral ...delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from Addgene...distributed in 2023 alone. You’ll also find a large number of requests for vectors in the “controls” category... usefulness of each viral prep in practice. The number of requests shows how many scientists believe a...

  11. The Challenges of Cell Culture

    Type
    Blog Post
    ...throw them away and thaw a new batch. The exact number of passages can vary between cells lines, as this... broadly recommended that the lower the passage number the better for experiments. This is to ensure the...and Working stocks of cell lines have the passage number recorded on the outside of the vial and in a cryostore...the front of the incubator containing the passage number, cell line, and date, which you complete each time...wishes to use these cells can see their passage number and key details at a glance. You should also monitor...

  12. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...Gibson assembly methods If you want to scale up the number of gRNAs in your plasmid, you’ll need to use some...to express 2-4 gRNAs, with four being the ideal number. First you generate four unique kanamycin-resistant...different destination vectors depending on the total number of gRNAs you wish to clone, from 2-7. For example...spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number). The 5’ most gRNA is cloned into the Cas9-containing...destination vectors are used depending upon the total number of gRNAs you wish to clone, so you don’t have to...

  13. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    ... to wasted time or retractions. Using a catalog number for identifying a reagent can be helpful, but it...online. Furthermore, if the link between the catalog number and reagent description is lost (e.g. a supplier...supplier is bought out by another company and catalog number changes) the original identifier loses its meaning...plasmids. All plasmids at Addgene have unique ID numbers associated with them, which serve as unique PIDs...

  14. Antibodies 101: Isotypes

    Type
    Blog Post
    ...five isotypes, such as the number and location of disulfide bonds, the number of constant domains, and ...heavy chain constant domains. These are typically numbered, i.e. IgG1, IgG2, etc. Let’s dive in and meet ...valency of two. There are four IgG subclasses, numbered based on their abundance in blood: IgG1, IgG2,...

  15. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... sensitive enough to detect gene copy number based on the number of fluorescent puncta observed. By labeling...approach, fluorescently labeled Cas9’s come with a number of limitations. Since color is specified by PAM...depending on the hairpin combination bringing the total number of individual colors to 6. A gRNA with all three...

  16. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ...increase the sensitivity of an assay by increasing the number of signaling molecules associated, indirectly, ...source and are readily available conjugated to a number of different signaling molecules. With the direct...of primary antibodies can increase the possible number of target proteins.   The exception to this rule...secondary antibody will be required regardless of the number of protein targets, simplifying multiplex assays...

  17. The time and cost required to make a plasmid

    Type
    Blog Post
    ...over a hundred. Of course, these were all 2018 numbers, and the intervening five years, inflation has ...experiments. This cost varies hugely based on a number of factors, but we can assume it’s at least equal...   Designing precise calculations for any of the numbers in this post is a mathematical story well beyond...large? Using Doench’s calculations and the 2018 numbers for consumable costs, our depositors have helped...

  18. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...carried on the transfer plasmid or quantifying the number of viral copies that have integrated into the target...measuring functional titer uses FACS to count the number of target cells that are positive for vector encoded...of lentiviral titration uses qPCR to measure the number of proviral copies that have integrated into a ...or the transgene itself are used to quantify the number of integration events. Another drawback to this...

  19. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...large number of inserts to assemble. You could also consider using Gateway if your insert number is a ...particular, another important consideration is: The number of inserts ...as some methods are more amenable...efficient; although efficiency often declines as the number of fragments you’re hoping to assemble increases...

  20. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...Additionally, CAPTURE 3C-based sequencing requires a large number of cells (~5x107). Together, these two requirements...in eukaryotic cells. Using this tag reduces the number of CAPTURE components that need to be delivered...BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs Number of loci targeted One Many Sample type(s) Cell...such as ChIA-PET or Hi-C, by increasing both the number of unique chromatin interactions identified and...
Showing: 41 - 60 of 415 results