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We narrowed to 798 results for: Cre

Showing: 301 - 320 of 798 results

  1. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ...needs to be compensated out of APC-Cy7. After increasing the compensation values for APC against APC-Cy7...to be shifted towards the x-axis, so you would increase the compensation value for APC. If the population... to be shifted away from the x-axis, you would decrease the compensation value.   Figure 1: Compensation...population needs to be shifted towards an axis, increase the compensation value, whereas if it the population...population needs to be shifted away from an axis, decrease the compensation value. If neither of these helps...

  2. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ...detectors for all fluorophores. This allows for the creation of much larger panels, as fluorophores that cannot...they don’t “wash out” the rest of the panel and create spreading error. Conversely, markers with low expression...should not be paired with these dyes, as this would create difficulty when trying to resolve those populations...signature upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical...spectral flow cytometry has the ability to generate incredible amounts of data from a single experiment. Although...

  3. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...amount of agarose/DMEM should be proportionately increased). For some cells, add a drop of serum to aid in...up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...either initially, or using secondary biological screens. For example, for cancer researchers, if growth...

  4. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ... to its final destination. As an example, most secreted proteins are produced in the endoplasmid reticulum...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ... they are finally degraded (2, 3). Once you've created your fusion protein, it's important to validate... to make any fluorescent protein constructs you create in your own work available to the Addgene community...Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka...

  5. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with a SmaI or XmaI restriction...duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary. To save time...mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial pellet...how iodixanol purifications works, shows how to create the iodixanol gradient layers, and has some great...master iodixanol AAV purification. If you’re new to creating gradients, practice making the iodixanol layers...

  6. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...suppression of background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion...obtained if any single component was omitted. Screening for successful Golden Gate Assembly All Golden..., Table 1). These improvements (5- to 12-fold increases in the number of transformants while maintaining...to faster assembly workflows that require less screening to confirm the accuracy of assembly. The fidelity...test the limits of DNA assembly approaches with increasingly complex experimental conditions. The ability...

  7. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ... aaRSs in these processes. To create the antibody toolbox, they screened antibody candidates using a synthetic...fourteen aminoacyl tRNA synthetases (aaRS) have been created, characterized, and validated by the Gräslund lab...cross-reactivity, antibody candidates underwent affinity screening to select for slow off-rate binders. The best ...pathways By Gabrielle Clouse Claudia Vickers’ lab has created a novel nanocompartment for transporting proteins...

  8. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... deletions or homology-directed repair (HDR) to create precise genome edits. But have you heard of another...depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh...PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for...PITCh-gRNA, and the locus-specifc gRNA. The PITCh-gRNA creates two DSBs in the PITCh plasmid to release the cassette... alter expression of the gene it follows. For increased versatility, it would be advantageous to adapt...

  9. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    Published
    Sept. 23, 2014, 3:05 p.m.
    ...Website: http://protocols.io/ Protocols.io is the creation of ZappyLabs. Currently in beta testing, the software...The site allows you to comment on protocols and create lists of your favorite protocols for future reference...of the early protocol sharing websites. It was created in 1999 and allows scientists to contribute protocols...web-based lab management tool that allows labs to create accounts in which their members can share protcols...your lab’s needs. At Addgene, we’ll continue to create new molecular biology educational resources, such...

  10. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ... or the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA...resistance gene. These reporters allow you to easily screen or select for cells that are expressing your gene...interest at high levels. Unlike vectors that express screenable or selectable markers from a unique promoter,...now download Addgene's new eBook! Our goal was to create a one-stop reference guide for plasmids. We've ...

  11. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...(CIB1) proteins. The goal of both groups was to create a system that would use light to turn on and off...domain to each of the Cas9 fragments, the authors created a photoactive Cas9 tool, as shown in the figure...binding and cleavage. This technique was applied to create photoswitchable spCas9, dCas9, and S. aureus Cas9...Gilgenast TG, Gong W, Beagan JA, Raj A, Phillips-Cremins JE (2019) LADL: light-activated dynamic looping...

  12. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins, making the purification...production of Fab fragments where variable chains are secreted into the periplasmic space to be oxidized and ...incubation time to maintain cell viability and increase protein production (Schwarz et. al., 2020). Harvest...   Figure 1: The steps of rAb production. Created with biorender.com   Another method of harvesting...contaminating bands or a few strong ones, consider increasing the number of washes performed during your harvest...

  13. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ... 2021). In fact, completion of Provost’s CURE increased students’ confidence in their research capabilities...projects have also been proposed as a mechanism for increasing diversity and inclusion in STEM disciplines (... Brownell, 2014; Duboue et al., 2022), as they increase student interest in scientific career paths while... Provost believes these networking experiences increase student enthusiasm, and research backs up that...undergraduate research experiences are a “CURE” that increases engagement by students and teachers. FEMS Microbiology...

  14. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...the templated DNA. Check out our blog post on increasing HDR efficiency to read more about optimizing ...pathways have been shown to be efficacious for increasing knock-in events (Yang et al., Int J Mol Sci)....culture media prior to and after Cas9 introduction increases HDR, just make sure to remove the inhibitors within...Early HR factors fused to Cas9 have been shown to increase HDR frequency by committing a break to performing...reading up on validating genome edits to find screening strategies for your experiment. Also learn about...

  15. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...large number of constructs for high-throughput screening of PPIs. For example, Bischof et al. (2018) generated...generated an open reading frame (ORF) library to screen PPIs among 450 transcription factors in live fruit...organisms lags behind. BiFC can also be used to screen for small-molecule inhibitors of a particular PPI...Here, one of the endogenous PIPs is expressed at increasing levels alongside your BiFC constructs. If your...co-expressing one of the endogenous PIPs should decrease the BiFC signal in a dose-dependent manner. This...

  16. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ...CRISPR, and the challenges of science becoming increasingly politicized. It would be nearly impossible for...all of these things contributed to the steadily increasing distrust the general public has in science and...self-promotion, and personality is playing an increasingly important role in science. The science produced...reserved Franklins and Prashers will find it increasingly difficult to compete (for funding, etc.) with...chapter on deep learning addresses the dilemma of creating a technology that could eventually replace scientists...

  17. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...newest line of defense against bacteria. Created with BioRender.com.   The problem of antibiotic...may also induce cell death. CRISPR also enables increased specificity - rather than targeting a process ...mouse skin colonization model, phagemid treatment decreased the proportion of virulent S. aureus from 50% ...death. Image from Beisel et al., licensed under a Creative Commons Attribution 4.0 International License....rendering the population sensitive to antibiotics. Created with BioRender.com. Phages could be used to...

  18. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    Published
    March 20, 2014, 3:04 p.m.
    ... shalt create community to help Involve family. If you don’t have local family to help, create “fake” ... on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work... to church, join a club, meet the neighbors and create a network of support to help with elder care, carpooling...partner – no kids. Get a massage – no guilt. Have recreational time with your friends – no spouses or kids....

  19. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...is possible with efficiency >70%. CRMAGE is an incredibly fast protocol, with only 5 hours incubation time...system that combines recombineering with CRISPR to create a system for scarless, iterative genome engineering... pathways within this genus, Huimin Zhao’s lab created two “pCRISPomyces” systems for use in Streptomyces...CRISPR in bacteria, they also led early efforts creating tools for CRISPRi and CRISPRa (Bikard et al., ... in Mycobacterial species, Sarah Fortune’s lab screened 11 other Cas9 orthologs to identify a more efficient... used to engineer the bee gut microbiome. They created a modular system using a broad host range plasmid...

  20. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...to solve this problem. This tool was initially created to support the CRISPR analysis needs of Synthego... sgRNAs. Since multiplexing is commonly used to create functional knockouts or large deletions, the ability...optimizing and no complicated steps to learn. For increased flexibility and scalability, the ICE software ...editing analysis Once the analysis is complete, a new screen appears with a graphical representation of the ...Data button on the bottom right of the analysis screen. ICE analysis details Contributions   The Contributions...
Showing: 301 - 320 of 798 results