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We narrowed to 856 results for: ELL

Showing: 481 - 500 of 856 results

  1. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...electrocompetent cells in which the phage recombination machinery is induced using L-arabinose. These cells can be...function of certain regions in a promoter or gene, as well as to study the effects of inactivating/activating...coli to undergo nick repair. This method can work well, but poor template digestion can result in a high...required in recombineering. REPLACR-mutagenesis is also well suited for insertions and deletions. The figure ...Enzymes Required No No Yes Yes Lab Made Competent Cells Required Yes No No No Comparison of REPLACR-mutagenesis...

  2. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    Published
    July 28, 2015, 2:30 p.m.
    ...also gives cells antibiotic resistance to neomycin so that you can select the population of cells with the...transcription factor. The FLAG antibody has been well characterized and the system allows you to add this...Only a small percentage of the proteins inside a cell have a commercial antibody against them. There’s...antibody. It’s an epitope not found in human or other cells. It was made artificially. That means you aren’t...lab already should be able to run this protocol as well. We would also like to put some social pressure ...

  3. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...we virtually always use every single well in a 96-well (or 384-well) plate! How much do repositories help...in this post is a mathematical story well beyond my ability to tell. But even from these rough estimates...depositors have helped us save the scientific community well over $80 million on plasmids alone. (And we expect...

  4. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ... be expressed in the same cell, but not necessarily at the same subcellular compartment as the opsin or...modulate the activity of neurons and other brain cells. Since the opsins or chemogenetic receptors used...neuropil conceals identification of opsin-expressing cells). For other things to keep in mind when making fluorescent...cerevisiae by use of an epitope addition method. Mol Cell Biol 8:2159–2165 . https://doi.org/10.1128/mcb.8.5.2159...Paré J, Bonaventura J, Richie CT, Michaelides M, Mueller SAL, Roseboom PH, Oler JA, Kalin NH, Hall RA, Smith...

  5. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...product, the transformation may not result in any cells, or all the colonies screened might not contain ...i.e. primer3) or provided by the major companies selling PCR reagents. Nowadays commercial polymerases are...verify the strain and the efficacy of your competent cells (when they get old, they lose competency). If all... look at your experimental design. Full lawn of cells. The antibiotic in your plate might not work. Check... the incubator. The small colonies are called satellite colonies and they form when all the antibiotic...acknowledged in hundreds of publications including Cell, Nature, and other journals.   Paolo Colombi is ...

  6. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...-HF1, & HypaCas9. The off-target problem It is well known that CRISPR/Cas9 genome editing can result... the EMX1 locus in human embryonic kidney (HEK) cells, 11 of these initial mutants retained the on-target...off-target activity at multiple genomic locations in HEK cells. Further testing of two of the mutants, SpCas9(K855A... showed that these mutants are not toxic to HEK cells and that similar mutations can improve the specificity... researchers to make precise edits in mammalian cells and may decrease worries about off target effects... Cas9 in complex with guide RNA and target DNA." Cell 156.5 (2014): 935-949. PubMed PMID: 24529477. PubMed...

  7. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...process will begin. Partnering with seqWell, we are using the plexWell technology to easily and quickly ...ever worked to sequence DNA on NGS platforms will tell you, while the process has become much more refined...assembly process. Again, thanks to our partners at seqWell we make use of a pipeline that takes the raw data...necessary adapter sequences for Illumina sequencing as well as unique barcodes per sample so we can assign sequencing...Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller I, Mitsopoulos A, Fan M (2020) A Novel Next-Generation...

  8. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    Published
    Sept. 13, 2016, 2:30 p.m.
    ...Communications, Harvard University Ray Howell -Founder of Howell Communications Philip Brenner, PhD - Assistant...scientific enterprise and the way it is funded have been well documented over the past 2 years (Alberts et al,... to shape the sector throughout their careers (McDowell et al, F1000Research, 2014; Dolan, F100Research...relationship, along with, of course, union dues. Time will tell the extent of the implications of this ruling and...27175853. Pubmed Central PMCID: PMC4866822. 3. McDowell, Gary S., et al. "Shaping the Future of Research...

  9. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ... acid (DAP), a component of the cell wall. Once in the recipient cell, the CRISPRi DNA (purple) flanked... is propagated during normal DNA replication and cell division. Mobile-CRISPRi does not disrupt gene function...mobilize these plasmids for transfer to recipient cells during a tri-parental mating scheme. Mobile-CRISPRi...bacteria associated with human disease (S. aureus, Klebsiella pneumoniae, and P. aeruginosa) in addition to...antibiotic trimethoprim. When Enterobacter aerogenes, Klebsiella pneumoniae, and P. aeruginosa (all opportunistic...

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

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...AmpliCan We chose to adapt Crispresso by Luca Pinello and Kendell Clement for batch analysis because of its...Leonetti Ryan Leenay Jason Li Max Haeussler Luca Pinello Kendell Clement Andy May Additional resources on the...members have taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To ...all the standard information that goes into a 96-well plate and comes out of it. Biotech companies are...

  11. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...there isn’t a template strand left behind that the cell can use to repair. Figure 3: General overview...Occasional cleaving of the target strand is not as well understood but likely occurs in similar regions....complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394.e3. https://doi.org/10.1016/j.molcel...formation by a type I-C CRISPR Cascade. Molecular Cell, 83(5), 746-758.e5. https://doi.org/10.1016/j.molcel...broad and unidirectional genome editing in human cells. Nature Communications, 10(1). https://doi.org/10.1038...CRISPR–Cas systems for genome engineering in human cells. Nature Biotechnology, 37(12), 1471–1477. https:...

  12. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ... make it difficult to properly gate your desired cell population. By compensating between fluorescent ...online manual for your cytometer should be able to tell you the laser configuration. In general, if two ...Figure 1A). Applying compensation means you are telling your cytometer to adjust the fluorescent signals... your fluorescent markers are staining for. If a cell is positive for both markers, it will show up as...Figure 3: Flow plot for a double positive sample. The cell population in the top right corner of the plot is...learning! Ashlyn has a research background in CD8+ T cells and cancer immunotherapies. She is currently a research...

  13. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ...different kinds of information on our plasmid pages? Well, to help make the most of these pages, we've written...Addgene’s plasmids is an indication of popularity. A yellow flame indicates that a plasmid has been ordered...process and these can be found in this location as well. These maps make it easy for scientists to see if...analyzed using Addgene’s Analyze Sequence tool. Excellent, how can I check if the plasmid will be within...resistance, and optimal growth temperature are listed, as well as the name of the strain.  Most Addgene plasmids...such as some of our viral vectors, in NEB Stable cells. Resource information: This section contains information...

  14. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...favorite parts for plant synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning (MoClo)...really easy to clone genes for expression in plant cells. Sylvester and I chose the 96 most useful things...used as we have tested them to show that they work well in plants and give similar stability to transcripts... parts available in your lab - if you get the 96-well kit - everything is so easy. We have undergrads ...a real person, so that students see academics as well-rounded human beings. It’s also a great communication... that want to have a big impact on the world and tell them to work in plant science. I think their chances...

  15. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...CRISPR complex much easier in various cells, including primary mouse cells. We believe that this system will...In 2014, you joined Addgene's Advisory Board, as well as the Editorial Board of Scientific Reports and...US, I very much enjoyed interactions with many excellent colleagues at NYU and visiting scientists, such...In addition, I've been very lucky to work with excellent colleagues in my lab. On the other hand, I feel...good for Japanese science. I really miss the intellectual environment in the US in this regard. In addition...

  16. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...occurs when bacterial cells regulate gene expression in response to changes in cell density. At the mixer...cohort of friends and colleagues had moved on as well, so I knew very few people. I didn’t really fit ...Singletons gained from spending time with a mentor, as well as having a cohort experiencing similar situations...introduced to several people by their mentor, as well as their bin-mates.  They liked us. They really ...Hendrickson, Jonathan Jacobs, Mo Kaze, Christina Kellogg, Heather Martinez, Pat Schloss, Tara Smith, Marc...

  17. Lab to Office Culture Shock

    Type
    Blog Post
    Published
    Feb. 5, 2019, 1:04 p.m.
    ... it, and many of my scientist colleagues have as well. Making the transition from the bench can be jarring...scientist going through even the worst experimental dry spells. Losing that sense of discovery can be hard since...place. The feeling of camaraderie one gets from a well-functioning lab team. Obviously not everyone gets...applying. I probably did not respond to her question as well then as I would now. I would have to repress laughter...the right set of tools to help you do your job as well as possible. It’s in their best interest to do so...the newer members of our Scientist team says, “a well-run meeting where everyone has a voice and the group...

  18. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    Published
    July 29, 2025, 1:15 p.m.
    ...editing reagents into cells. Viral vectors, because of their natural ability to infect cells, have emerged as...biologist, academic, and researcher. Among the most well-known CRISPR-associated nucleases are Cas9 and Cas12a...which poses a challenge for efficient delivery into cells, particularly via viral vectors. While viral delivery...for gene editing and allows targeting of specific cell types, it has a limited capacity to carry large ...biolistic, or PEG-mediated transformation into plant cells. Conclusion & Future Directions TnpB represents...

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

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ...recommend transforming into chemically competent cells in a 96-well PCR plate. Always confirm your results Congratulations... sequences of the parts and acceptor vectors, as well as the restriction enzymes, and it will create the...transfer and the restriction–ligation reactions in a 96-well (or higher!) format. Liquid handling robots are ...Dr. Rob Hurt for high-throughput minipreps in 96-well plates. Plate wisely You will likely see fewer numbers... this, you can increase the volume of competent cells you plate after each transformation and/or use different...

  20. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ... edit A → Y (Y = T or C) in 67.17% of screening cells (Tong, Wang, et al., 2023). Based on this promising...the base editor. Black arrows indicate natural cellular processes. Created with BioRender.com. The...Cas9 nicks the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template... efficient A•T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4), 638–650...programmable T-to-G and T-to-C base editing. Molecular Cell, 84(7), 1257-1270.e6. https://doi.org/10.1016/j....efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547. https:...
Showing: 481 - 500 of 856 results