We narrowed to 795 results for: Kin
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TypeBlog PostPublishedNov. 11, 2020, 2:23 p.m....replication machinery room to come in and get busy making copies. Types of origins of replication ...compatibility with one or more other plasmids. Generally speaking, plasmids with the same ori are incompatible because...chloramphenicol – make sure your growth conditions aren't working against you! The culture inoculum Freshly ...
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Safe Port in a Storm...How Addgene is Weathering the Pandemic
TypeBlog PostPublishedSept. 17, 2020, 1:15 p.m....Addgene started dealing with this new reality of working remotely, scientists around the world mobilized...took to make all this happen with 80% of the team working remotely. We’re extremely proud of the work we’... maintain our workforce for when we need to be working at full capacity to serve the scientific community... -
Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative
TypeBlog PostPublishedJune 2, 2020, 1:15 p.m....from Zhu et al., 2019. Wensheng Wei's lab at Peking University has developed a new CRISPR screening...injection of precise titrations of AAV or heat-shocking the organism. The SPARC system uses competitive...fluorescing green. Image from Shin et al., 2019. Looking for an alternative to intercalating fluorescent... -
Meet the New Addgene Board Members!
TypeBlog PostPublishedApril 28, 2020, 1:14 p.m....around the world who would otherwise have difficulty making or getting these reagents is profound.” Frank Stegmeier...Library, a library that “enables the high-resolution tracking of more than 1 million cancer cells under drug...became passionate about virology,” he says. He was working in an HIV lab and based on that experience, soon... -
R Bodies: Membrane-Rupturing Microscopic Tools
TypeBlog PostPublishedApril 14, 2016, 2:30 p.m....biological motion (whether endocytosis, vesicle trafficking, or muscle contractions) are produced by orchestrated...the ciliate. It’s thought that they function as a kind of armor to protect their host from predation (Petroni... -
Components of CRISPR/Cas9
TypeBlog PostPublishedFeb. 2, 2016, 5 p.m....Cas9, and the tracrRNA which acts as a scaffold linking the crRNA to Cas9 and facilitates processing of...different components of CRISPR, It's time to start thinking about how you can used them in your experiments...experiments. Use the following flow chart to get you thinking about the types of experiments you're planning on... -
The Importance of High Titer for AAV Transductions
TypeBlog PostPublishedMarch 1, 2018, 2:11 p.m....Check out this post for a refresher on AAV titers Making contact with a cell Although an individual AAV ... this type of vector to such a high titer when looking for effective gene expression. How do you achieve... Hanley is a Research Team Lab Tech at Addgene working on methods to improve AAV production. References... -
The Power Behind NGS Plasmid Validation: seqWell
TypeBlog PostPublishedApril 19, 2017, 3:25 p.m....“reading”) of DNA, have changed our collective thinking about the feasible size and scope of projects ...library prep. As part of our mission, we’ve been working with Addgene to develop and apply our plexWell™...read level, and because custom primers or "primer-walking" reactions are completely unnecessary for sequencing... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog PostPublishedSept. 3, 2019, 1:15 p.m....contributed by Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells...These loops not only provide a 3-D structure for packing genomic DNA into nuclei, but are also thought to... -
Summer SciComm Series: A PhD in Science Communication
TypeBlog PostPublishedAug. 23, 2022, 1:15 p.m.... apart sources of information they don’t like, looking for reasons to dismiss them. There’s evidence to...Zealand’s multiple Covid-19 lockdowns, I held mask-making workshops in my community. I brought along printed... to better meet the needs of the people we are talking to. Emma Harcourt is a PhD student in the Department... -
Hot Plasmids - January 2023
TypeBlog PostPublishedJan. 31, 2023, 2:15 p.m....hot plasmids articles. Think we missed something smokin'? You can pitch a Hot Plasmids section (antibodies...heatshock promoter) drives expression of the tumor-seeking CAR (chimeric antigen receptor), whereas a ubiquitous... variants with similar capabilities. If you’re looking for a super stable FP for your next microscopy ... -
Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis
TypeBlog PostPublishedAug. 28, 2014, 3:34 p.m....idea to take some time to confirm that you are working with the correct construct, and verify that the...Consult the manufacturer’s manual for the optimal working conditions for each enzyme. Watch out for methylation...run control uncut DNA to ensure your enzymes are working. When uncut plasmid DNA is isolated and run on ... -
Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering
TypeBlog PostPublishedMay 3, 2018, 1:48 p.m....These enzymes also do not require a protospacer flanking sequence, so you can target virtually any RNA ...Cas13d enzymes have a median length of ~930aa - making them 17-26% smaller than other Cas13s and a whopping...and we at Addgene appreciate our depositors for making their tools available to the broader community.... -
A Practical Approach to Choosing the B(right)est Fluorescent Protein
TypeBlog PostPublishedJune 1, 2017, 2:30 p.m....reference protein) under identical conditions, a ranking of the practical brightness can be made (Goedhart...repeating this with another fluorescent protein, a ranking of practical brightness can be made. This strategy...their practical brightness in a system closely mimicking the future application. In figure 2 we provide... -
Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection
TypeBlog PostPublishedMay 11, 2017, 2:30 p.m....high-throughput manner and therefore push us closer to making cancer precision medicine a reality. Challenges...Since time is of the essence in clinical settings, looking ahead, we’re starting to build saturating libraries... Institute of Harvard and MIT and is focused on making precision functional genomics a reality for future... -
15 Years of Addgene: The Top 15 Plasmids
TypeBlog PostPublishedJan. 8, 2019, 1:54 p.m....great? It produces a ~10 fold higher titer virus, making it ideal for CRISPR pooled library screening experiments...V2.0 was made available through Addgene in 2015, making this the most recently available plasmid in the... thank all of our depositors and requestors for making their reagents available and for supporting scientific... -
AAV Q&A with Tim Miles
TypeBlog PostPublishedMay 17, 2022, 1:15 p.m....question that didn’t get answered live - but he kindly took some time to address all your unanswered questions...astrocyte specificity and would like to get it working in microglia as well. A: We are not aware of any... tail vein injections but would like to avoid "leaking" into the brain, would you recommend using the ... -
Viral Vectors 101: The AAV Retrograde Serotype
TypeBlog PostPublishedApril 27, 2023, 1:15 p.m....can begin to understand how the brain works. It’s kind of like “the head bone’s connected to the neck bone... to the rest of the body. Therefore, by simply looking at how neurons are physically connected we can ...efficiently deliver genetic information or tools, making them useful tracers, but finding a virus that works... -
Viral Vectors 101: Viral Vector Elements
TypeBlog PostPublishedJuly 25, 2023, 1 p.m....think. So, if you’ve ever wondered what goes into ‘making virus’ or what gag and pol stand for, we demystify...recombinant adenovirus is missing the endogenous E1 gene, making it replication deficient. This is typically supplied...discussed are compiled for reference below. Happy virus making! References and Resources Kulanayake, S., Tikoo... -
Antibodies 101: Monoclonal Antibodies
TypeBlog PostPublishedJune 29, 2021, 1:15 p.m...., you may be curious about all of the different kinds of antibodies that are available. One common type...References Bradbury ARM, Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B,...TA, Humphries MJ (2016) Ligand-induced Epitope Masking. Journal of Biological Chemistry 291:20993–21007... -
Behind the scenes: Addgene’s new search engine and more
TypeBlog PostPublishedDec. 12, 2019, 2:22 p.m....— by directly typing terms in a search bar, by ticking some boxes, by selecting a suggestion — how should...resources. Filters We identified the most useful kinds of information per resource category to expose as...our search indices in order to support different kinds of matches: exact words, partial words, phrases,... -
Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens
TypeBlog PostPublishedFeb. 24, 2015, 7:50 p.m.... it’s working. You can only do that screen with a system you are really confident about working for most...made standard protocols available and they are working to further optimize those now. They are also developing... -
3 Challenges in Plant Synthetic Biology
TypeBlog PostPublishedJuly 22, 2014, 5:46 p.m....better fuels, feeds, foods, and fibers. And because working with plants can be challenging, there are a lot...construct our color changing flower we are carefully making our way through the IP landscape, using plant parts...novel pieces where applicable. We will also be working with open source, transformation technologies like... -
Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur
TypeBlog PostPublishedJuly 10, 2014, 7:17 p.m.... and devoted 100% of his energy and effort into making the startup self-sustainable. Within a year from...all safety nets.” Raising seed money. “Before seeking funding, ask yourself if you really want to do ...Co-founder and formerly VP Operations of Sample6) for taking the time to speak with us about the biotech startup... -
Choosing the B(right)est Fluorescent Protein: Aggregation Tendency
TypeBlog PostPublishedJune 15, 2017, 2:30 p.m....in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...for individual cells, and the number of 'normal' looking cells can be counted. This assay will provide information...on the dimerization tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not ... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog PostPublishedMarch 19, 2019, 1:31 p.m....inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument for controlling ...and systematically mutated it. Using a robotic picking approach to select cells with variant characteristics...al, Nat Chem Biol 2018. PubMed PMID:29483642. Making protein-protein interaction studies easier with... -
Antibodies 101: Fab Fragments
TypeBlog PostPublishedSept. 5, 2024, 1:15 p.m....this point, you may be asking yourself, what type of Fab fragment are we talking about here? Many of the...fragment. But this bivalency may cause problems for blocking purposes, since the fragment could bind not only... -
Antibodies 101: Affinity Reagents
TypeBlog PostPublishedApril 14, 2022, 1:15 p.m....someone speaks about an affinity reagent, they’re talking about a standard antibody: An ~150kD protein with...nanobodies, are derived from a camelid antibody lacking a light chain, called a heavy chain antibody (Muyldermans...clear: it’s an exciting time to be a scientist looking for that perfect fit! References and Resources... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....between them, could affect the properties or trafficking of the fusion protein (Kimple et al., 2013). ...opsin-expressing cells). For other things to keep in mind when making fluorescent protein fusions, check out Joachim ...Fluorescent Proteins 101: GFP Fusion Proteins – Making the Right Connection. Thank you to our guest... -
Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux
TypeBlog PostPublishedSept. 13, 2016, 2:30 p.m....Advancement of Science, Rush Holt, followed by a networking reception. Saturday’s workshop will be an advocacy...laid out by a 2014 National Academy of Sciences working group (National Academy of Sciences. “The Postdoctoral... Advancement of Science, Rush Holt, PhD and a networking reception. Saturday’s workshop will progress ... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m.... me troubleshoot some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements...cells for infection If your transductions aren’t working, you can check if virus is being generated by your...template. Regardless of the type of virus you’re working with, there’s a lot that goes into virus design... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m....The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...amount of virus you actually have in your prep. Taking time to properly titer your virus will not only...original plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will ... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m....density: particle concentration is measured by looking at absorbance at 260nm using UV/Vis spectrophotometry.... The purity of the sample can be estimated by looking at the ration of DNA (260nm) versus protein (280nm...calculated in terms of plaque forming units (pfu), taking initial vector dilution into account. You'... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog PostPublishedJune 27, 2017, 1:03 p.m....or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, ...articles describing the FRET tool that you are looking for and check our curated list of biosensors too... Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking the ruler to the jungle: Single-molecule FRET for... -
Plasmids for Endogenous Gene Tagging in Human Cells
TypeBlog PostPublishedApril 6, 2017, 1:02 p.m....alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope...whole segment—a GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into...and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing the tag... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m....Scientists looking for more compact genome editors due to delivery constraints and scientists seeking a wider... prep in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....laboratory of Dr. Moshe Szyf at McGill University, working to develop CRISPR-based tools for epigenetic editing...Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun...Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013) TET1 Controls CNS 5... -
Hot Plasmids February 2024
TypeBlog PostPublishedFeb. 20, 2024, 2:45 p.m....10.1038/s41587-023-01756-1. PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants ...with some applications. Several labs have been working to produce monomeric versions that break up the...was delivered to brain endothelial cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when... -
7 Simple Strategies to Resolve Conflicts with Difficult Supervisors
TypeBlog PostPublishedJan. 21, 2015, 1:30 p.m.... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...explain how it is interfering with your work. Avoid talking about your emotions, such as frustration or anger...