We narrowed to 1,026 results for: ELL
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TypeBlog Post...community. Addgene's AAV preparations have worked well in my lab, including in applications where we have... can be used to deliver fluorescent proteins to cells as determined by the serotype. You can use these...determine if your AAVs are getting to the right cells using your particular delivery technique and conditions...expanding our viral vector educational resources as well, with both posts on the blog and pages on the Addgene...website you’ll find our updated viral vector pages as well as expanded virus protocols, information on how ...specific material page and includes western blots as well as other expression data. If you have any questions...
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Don’t Wait To Launch Your Health Science Startup
TypeBlog Post...would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am lucky...your life making connections and finding mentors -- well, it's time you call upon these people for guidance...Don’t go it alone. You’ll find that no matter how well you know your idea, you will rarely succeed in launching...partners. You need a dedicated team that you know well and trust. This will allow you to discuss each ...two close friends, Sreyan Chowdhury and Clinton Buell, who have very diverse educational backgrounds (... -
Advice for Moving Into Sales After Your Science Postdoc
TypeBlog Post...Fhearraigh, PhD, of www.phdadvice.com. I sold out, well in part that’s what leaving academia feels like ...would continue on with my academic dream of studying cell division. However, after 6 years in the business...products, so you have some of the best ideas on how to sell these products to other scientists. Tailoring my...introduction to the financial world. It teaches you about selling products which is an important part of 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 Post...an awake monkey is different from a cell in a dish. I always tell people, 'Be driven by your science.'...for neuroscience back in 2005, both activators as well as silencers of neural activity. Since then, there...to Addgene in a matter of days. Addgene: Can you tell us a little more about this new two-color tool and...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 ... -
Revamp Your Lab Meetings With Creative Virtual Collaboration
TypeBlog Post...blogger Matteo Tardelli! Matteo Tardelli, PhD is Postdoctoral Scientist at Weill Cornell Medicine in NYC...was contributed by Matteo Tardelli, a postdoctoral scientist at Weill Cornell Medicine. Discussing data...Hamilton DP, Denfeld BA, de Eyto E, Hampton SE, Keller PS, Sharma S, Lewis ASL, Weyhenmeyer GA, O’Reilly... -
With Our New Viral Service, We're Taking CRISPR Further!
TypeBlog Post...of viruses used to deliver CRISPR components to cells or organisms under study. In the past we’ve left... they want to do is to quickly generate a single cell line and proceed to use it for experiments.” - ...screen for genes involved in a wide variety of cellular processes. While these are widely requested and...representation in the viral prep, infect target cells, and, finally, conduct the screen. To help alleviate...of this labor for our users, we now provide the Brunello (targeting human genes) and Brie (targeting mouse...deliver gRNA and therefore, must be used to infect a cell line already expressing Cas9. As John Doench (Broad... -
The Crispr Babies Saga Shows the Need for Action, Not More Delays
TypeBlog Post...now. Many thanks to our guest blogger, Kelly Hills! Kelly Hills is a founding bioethicist of the consulting...This post was contributed by Kelly Hills, founding bioethicist of Rogue Bioethics. On November 25, the... the MIT Technology Review dropped a bombshell report. A scientist working in China was using CRISPR/Cas9...separate things happened nearly simultaneously: The Wellcome Trust funded a center for mitochondrial research...consulting firm Rogue Bioethics. Kelly utilizes her firsthand experience in the software industry to inform... -
Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish
TypeBlog Post...multispecies biofilm formed by Pseudomonas spp. and Klebsiella spp. Dr. Devarshi Gajjar, The M. S. University...research focuses on a collection of pathogenic Klebsiella and Pseudomonas isolates causing urinary tract...unknown regulatory regions in Pseudomonas and Klebsiella spp. The effects of these deletions on biofilm...major pathophysiological complications of diabetes mellitus. Recombinant human proteins that play a role in... Researcher who investigates the molecular and cellular etiopathomechanisms of ischemic stroke, the most... reduced blood flow. Novel treatment options, as well as non-pharmacological solutions to modify the course... -
Plasmids 101: Gram Negative and Positive Bacteria
TypeBlog Post...negative bacteria is due to differences in cell wall composition. Cell wall structure Gram positive bacteria...used in the staining process bleached another, Klebsiella pneumoniae. After realizing this was a repeatable...membrane. Common gram negative bacteria include salmonella and E. coli. Because of their thicker peptidoglycan...complex within the peptidoglycan layer, staining the cell purple. When the alcohol is added, it dehydrates...Silhavy, TJ., Kahne, D. & Walker S. The Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414... -
DAP arrays for multiplex gene editing
TypeBlog Post...different types of human cells, including T cells and hematopoietic stem and progenitor cells. Now with the DAP...to enable multiplex gene editing in a variety of cells and organisms. However, lengthy genetic payloads... over 50% averaging editing efficiency in human cells. MBE with DAP array also exhibited reduced Cas9-... editing, and nuclease editing, in virtually any cell type or organisms of interest with only minimal ... -
Optimizing Donor DNA for Enhanced CRISPR Genome Editing
TypeBlog Post... between Cas9 (which causes the breaks) and host cell DNA repair factors (which repair those breaks) makes...example, correcting the allele that causes sickle cell anemia. However, HDR occurs much less frequently...a structure could drive sequence replacement in cells. Targeting three dCas9 molecules to a precisely ...functional Cas9 protein and the targeting guide RNA into cells by electroporation. Results from our lab and others... HDR. The detailed protocol for nucleofection as well as the sequences of our validated editing reagents...Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins... -
Viral Vectors 101: The AAV Retrograde Serotype
TypeBlog Post... into the cell. This means that not all neurons and pathways will be transduced equally well by AAV-retrograde...they either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...them useful tracers, but finding a virus that works well for retrograde tracing can be difficult. The in ... Data Hub!) The cortical pathways to the IC were well transduced by AAVrg while other pathways such as...chemogenetic vector directly into the region where the cell bodies of the projection neurons reside. Impact... -
Developing a cold tolerant E. coli using specialized chaperones
TypeBlog Post...government agencies, active researchers, as well as fellow iGEM teams. We were the proud hosts for the...industrial or research purposes requires stable host cells (the simplest example would be a transformed mesophilic...successful) as host systems since their biology is well studied, and they are convenient to maintain under..., since mesophilic chaperones would not function well under suboptimal conditions. This means recombinant...CRISPR-Cas mediated editing for psychrophilic systems as well as engineering and regulating psychrophilic recombinant... -
Plasmids 101: Inducible Promoters
TypeBlog Post...stimulate AraC binding to I1 and I2 sites. Supplementing cell growth media with glucose decreases cAMP and represses...Steroid-regulated LexA promoter XVE (synthetic) pFZ19 (yellow flame) Temperature inducible promoters Temperature...gene transcription to take place. Addgene also has yellow flame light-regulated two component systems designed... Because transcription machinery differs between cell types or organisms, promoters are similarly variable...variable. Bacterial promoters only work in prokaryotic cells and typically only in the same or closely related... were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc.) require unique... -
AAV Q&A with Tim Miles
TypeBlog Post...address all your unanswered questions via text! (well, maybe not all of them. We still haven’t gotten ...and would like to get it working in microglia as well. A: We are not aware of any available AAV for efficiently...co-delivering different AAV particles into the same cells in vivo? A: This will be highly dependent on vector...particularly one that can target the epithelial cells in the blood-brain barrier? A: For liver, we recommend...worth investigating for both Q6 & Q7.For epithelial cells of the blood brain barrier we would currently recommend...and to all our participants for their time and excellent questions. Additional Resources on the Addgene... -
REPLACR Mutagenesis: Replacing In Vitro Recombination Methods
TypeBlog Post...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... -
Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog Post...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 ... -
The time and cost required to make a plasmid
TypeBlog Post...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... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog Post... 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... -
Troubleshooting Your Plasmid Cloning Experiment
TypeBlog Post...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 ...