We narrowed to 664 results for: cell
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TypeBlog PostPublishedFeb. 18, 2016, 3:42 p.m....use electro-competent cells instead of the more common chemically-competent cells. You should perform, ... manufacturer’s instructions for your competent cells. For most standard cloning, you can transform 1-...1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using much less total...colonies, you might want to use higher competency cells. Additionally, if your final product is going to...
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Which Fluorescence Microscopy Technique is Best for Me?
TypeBlog PostPublishedOct. 10, 2017, 1:57 p.m....high light doses to the cells. Thin static samples Ex: Fixed monolayers of cells or thin (<15 μm) tissue...1). Thin dynamic samples Ex: monolayers of live cells with fluorescently labelled, motile structures These... 10s of nanometers. Thick dynamic samples Ex: 3D cell cultures, small model embryos >15 μm Thsee are some...Total internal reflection fluorescence microscopy in cell biology. Methods Enzymol 361, 1-33 (2003). PubMed...Richardson DS, Lichtman JW. Clarifying Tissue Clearing. Cell 162, 246-257 (2015). PubMed PMID: 26186186. Additional...Biological Imaging and a Lecturer on Molecular and Cellular Biology at Harvard University. No matter whether...Biological Imaging and a Lecturer on Molecular and Cellular Biology at Harvard University. Additional ... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m.... in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct NGG PAM NGA PAM NGC PAM NGT... percent editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages... -
Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria
TypeBlog PostPublishedApril 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... -
Antibodies 101: Flow Compensation
TypeBlog PostPublishedMay 16, 2024, 1:15 p.m.... make it difficult to properly gate your desired cell population. By compensating between fluorescent ... 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... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog PostPublishedOct. 26, 2017, 1:59 p.m.... Typically, Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation...DNA. This bias can be counteracted by growing the cells on solid plates, where each transformant forms its...own colony, uninfluenced by the growth of other cells. For libraries where the plasmids are all very similar... -
Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors
TypeBlog PostPublishedJuly 27, 2017, 1:11 p.m....reliably detect HMOs. Fatima’s first design is a whole cell biosensor with two components - an enzyme that breaks...binding results in an increase in fluorescence of cells that actively produce the HMOs. Fatima has demonstrated...out Fatima's recent publication on her HMO work in Cell Chemical Biology! Additional Resources on the Addgene...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their...proposes using this sensor to detect HMOs found on cellular surfaces. After incubating the sensor with bacteria... -
CASTing Off for New Shores in Human Genome Editing
TypeBlog PostPublishedJune 10, 2025, 1:15 p.m....they were barely effective at editing eukaryotic cells — at least, until they met the directed evolution...components. It drove 19% integration in HEK293T cells (more than 500 times better than the precursor it...R. (2025). Programmable gene insertion in human cells with a laboratory-evolved CRISPR-associated transposase... -
Choosing Your Fluorescent Proteins for Multi-Color Imaging
TypeBlog PostPublishedOct. 9, 2014, 3 p.m....to supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved...Imaging Center at UCSF. A common requirement for live cell imaging experiments is the ability to follow multiple... -
Advice for Moving Into Sales After Your Science Postdoc
TypeBlog PostPublishedAug. 21, 2014, 3:15 p.m....would continue on with my academic dream of studying cell division. However, after 6 years in the business...Cambridge University where he studied mechanisms of cell division. Recently Seán got the entrepreneurial ...information on ELISA troubleshooting, ELISA kits and cell death assays. You can find Seán on twitter @Macfhearraigh... -
Interview: Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering
TypeBlog PostPublishedMarch 4, 2014, 4:49 p.m....and see how different parts of the brain or the cells within them work together or perhaps compete against...two-photon properties, so you could activate single cells in arbitrary patterns in the brain. Addgene: Do ... different – an awake monkey is different from a cell in a dish. I always tell people, 'Be driven by your... -
Save Time with Transient Plant Leaf Transformations
TypeBlog PostPublishedJuly 25, 2019, 12:03 p.m....roots, as functional chloroplasts and leaf specific cell types are missing in roots. The development of transgenic...transformation” usual means that neither a stable cell line or plant is regenerated from the transformed...with the gene of interest is transferred into the cells. But for C. roseus, Agrobacterium infiltration into...Intrinsic GUS-like activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. ...fused to a protein of interest to elucidate the subcellular localization of your protein of interest. Find... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m....editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652.e29. https...Guide RNA and Target DNA. Cell, 156(5), 935–949. https://doi.org/10.1016/j.cell.2014.02.001 Yan, J., Oyler-Castrillo...type of edit, genomic context, delivery strategy, cell type, and goal of the experiment. Now, onto the ...nicking of the unedited strand and encourage the cell to use the edited strand as a template during DNA...also allows certain unintended edits to escape cellular surveillance. To limit unintended incorporation...https://doi.org/10.1016/j.cell.2021.09.018 Nelson, J. W., Randolph, P. B., Shen, S. P., Everette, K. A... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....machinery, and could thus be used in non-dividing cells. Cas13 enzymes also don’t require a PAM sequence... a maximum of 28% editing efficiency in HEK293T cells, as assessed using RNA-seq. Since the REPAIRv1 machinery...including reversibility and use in non-dividing cells. With the expanded targeting ability using both ...Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes." Mol Cell. 66(3) (2017):373-83. PubMed PMID: 28475872. Find... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....What works well in one cell or neuron type may not work well for a neighboring cell type, even within the... conditions (3,5,8), injection titers (1,2), and cell types used for packaging viral particles (6), can... vector performance in various brain regions and cell types. Power in numbers A Pubmed search for “AAV...options while AAV8 (82%) and AAV9 (84%) were both excellent at infecting our target neurons (Figure 1). However... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog PostPublishedJuly 12, 2021, 1:15 p.m.... investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids express these...for SARS-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their search, they found...packaging) and contain an IRES-Puro marker for stable cell clone selection. For more information on these plasmids...Bajic G, Harrison SC, Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-...depends on its spike protein to bind ACE2, the host cellular receptor. To more easily study the spike variants... -
DNA Purification Without a Kit
TypeBlog PostPublishedFeb. 11, 2020, 2:15 p.m....broken out of cells with a solution of salts, detergent, and ball bearings. Then a cellulose dipstick is... kits and typically follow the same principles: cells are lysed, RNAs and proteins are removed to leave... plant, microbe, blood, or mammalian cells or tissue Product: total nucleic acids While glass or silica...and RNA from plants, microbes, blood or mammalian cells in 30 seconds or less without using a pipette. ... could be digested with restriction enzymes. Cellulose nucleic acid purification Starting material: plant...silica are commonly used to bind and purify DNA, cellulose also efficiently binds, or at least entraps nucleic... absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids, so placing the... -
Simplify Cloning with in vivo Assembly
TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m....transformation efficiency of the strain: when asking the cell to perform multiple recombination events, all linear...linear fragments need to get into the same cell. For simple cloning requiring just one or two recombination...recombination events, homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications...modifications will require ultracompetent (109 CFU/μg) cells. Primer design Using specific primer design (Figure... -
Tips from the Repository Trenches: Using Barcodes to Track Samples
TypeBlog PostPublishedOct. 6, 2015, 2:30 p.m....using an electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...concentration or need to be transformed into different cells than were originally requested. Most of our samples... Start by scanning the sample barcodes into the cells in the same order that they exist within the rack...storage process - from deposited DNA to transformed cells and finally to bacterial glycerol stocks that are... -
Plasmids 101: Protein tags
TypeBlog PostPublishedDec. 11, 2014, 4:26 p.m....not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...