Skip to main content
Addgene
Open Menu

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

Information

In Category

We narrowed to 420 results for: Numb

Showing: 41 - 60 of 420 results

  1. 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...

  2. 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...

  3. 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...

  4. 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 ...

  5. 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...

  6. 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...

  7. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...assembly levels, you will be using an increasing number of parts in your one-pot one-step reactions and...efficiency, as concentration does not equal the number of DNA molecules. A solution of a longer DNA fragment...to perform your assemblies, but you have a high number to get through. It’s time to think big and perform...experimental costs. If you are planning to perform a high number of assemblies, you will also likely need to miniprep...plates. Plate wisely You will likely see fewer numbers of colonies (and therefore positive colonies) after...

  8. 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...

  9. 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...

  10. 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...

  11. 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,...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...particularly useful when trying to screen a limited number of cells like primary cells or in vivo. Recently...performance, an important criterion for probing a limited number of cells. Get the Brunello CRISPRko sgRNA library...essential for viability. In addition, high copy number genes may mimic viability genes as they are subject... while mitigating toxicity induced by high copy number dsDNA cutting. A Dolcetto library containing only...

  19. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...accessibility varied country to country.   Figure 1: Number of countries Addgene distributes to per year, 2005... 2005-2023. We identified and offered a number of solutions that increased accessibility, such as using...ice if the package stays in transit for a certain number of days and tracking average transit time for each...in a specific country can quickly reduce a large number of barriers. Though we haven’t solved every shipping...

  20. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ... system.  As the number of CRISPR reagents continues to grow, so too will the number of reagents available... cannot be modified using wild-type SpCas9. The number of off-target cleavage events for the VQR and VRER... and will continue to dramatically increase the number of CRISPR applications. The first non-Cas9 CRISPR...
Showing: 41 - 60 of 420 results