We narrowed to 712 results for: STO
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TypeBlog Post...for drafting my stories. In total, I prepare 5-6 stories for an interview, since one story usually has more...Instead, I preferred to prepare stories to answer interview questions. Story telling shares many of the same...of the theme of my interview stories as the characteristic which the story demonstrates I possess. Bullet...helpful for my interviews in industry. Prepare stories to tell in your interview The STAR interview method...answer: a setting, a conflict, and a resolution. Stories usually have a theme too, which is something not...of questions. Writing a rough outline of these stories helped me remember them for the interview. Prepare...problem, or making a key decision. Even deciding to stop pursuing a particular line of research is a deliverable...
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5 Tips for Troubleshooting Viral Transductions
TypeBlog Post... researchers prefer to store freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. ... in a given viral stock will give you a sense of the infectivity of your viral stock, save you time, and... compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles. Depending...these can be increased by concentrating a viral stock. Different titering methods can reveal differences... your virus by ultracentrifugation of the viral stock, followed by resuspension of the collected particles...highly sensitive to freeze-thaw cycles and should be stored in single-use aliquots. Never use a polybrene solution...increase transduction efficiency by 1.5-fold (Stockschläder et al., 1999). 5. Check your packaging cells... -
10 Basic tips for mammalian cell culture
TypeBlog Post... them, and store all but one in liquid nitrogen. The frozen vials make up your master stock, while you...be used as your working stock. This way you can go through your working stock vial by vial before thawing...another vial of the master stock. This ensures that you have many cells stored in their original condition...Keep the hood clean Hoods should not be used as storage cabinets. Clutter not only makes it difficult to... make things more manageable, but it also keeps stock solutions from getting contaminated. Make sure cells...like their space. If they reach confluence, they stop growing (contact inhibition) and it takes them time...condition and can replace your working stock if it gets too old or contaminated. This also ensures that anyone... -
Addgene's Tips for Plasmid Quality Control
TypeBlog Post...glycerol stock production and storage, we grow a little extra and transfer 100 µl of glycerol stock to each...that we are sequencing from the exact same stock that we store and distribute from. Of course, your culture...software such as Primer3 to help you design your custom primers. Remember, when designing a sequencing ... of our Sanger sequencing is done from glycerol stocks (overnight cultures with 15% glycerol), as opposed... culture. Sequencing directly from the glycerol stock saves us the step of purifying the DNA and gives...96-well PCR plates are prepared with the primer stocks for forward and reverse reactions, respectively...equipped to perform Sanger sequencing from glycerol stocks, so make sure to select the appropriate service... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog Post...field in which covalent modifications to DNA or histones, the proteins that help package DNA, were associated...modifications, researchers used blunt tools like histone deacetylases, but targeted epigenetic modification...researchers additionally fused a TAL effector to LSD1 histone demethylase to demethylate enhancer regions (Mendenhall...regulate transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and... acetyltransferase increases levels of H3K27ac histone modification at specified loci. Charles Gersbach... Figure 1: dCas9-p300 adds H3K27ac marks to histones in promoter and enhancer regions. These marks ...and repair pathways work downstream of Tet1 to restore the DNA after cytosine removal. Recently, Albert... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog Post...bound to modified proteins called histones. We can detect modified histones with “GPS for proteins,” ChIP-... of modified histones, a genomic region that has mono-methylated and acetylated histones, but not tri-...map with three coordinates: chromosome, start, and stop. For instance (chr3, 1, 10) indicates a stretch ...Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome ...tri-methylated histones, functions as an enhancer. We can therefore denote enhancers as follows: (mono_methylation...of epigenomics: H3 – one of NANOG’s associated histone proteins K4 and K27 – locations of the amino ...usually housed in a single flask. Through a bit of stochastic magic and Poisson statistics, each sub-population... -
27 Hot Plasmids from 2016
TypeBlog Post... are provided in both open (no stop codon) and closed (including a stop codon) formats. The collection..., et al. Cell Rep. 2015. PubMed PMID: 26190106 PRESTO-TANGO: An open-source resource for drug discovery...receptors. Bryan Roth's lab has a newly available PRESTO-TANGO kit which allows researchers to interrogate...and Screening via Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved ...discovery. Researchers can use plasmids from the PRESTO-TANGO kit directly for drug screening or easily...Luckily, researchers from Douglas Densmore’s lab at Boston University have greatly simplified the process ...CIDAR MoClo Parts Kit are available as glycerol stocks in a single plate with every plasmid or as individual... -
Getting the Most from Your Lentiviral Transduction
TypeBlog Post...thawing a fresh vial. In many labs, the history of the cells in storage may be a bit dubious due to lack of... especially when the lentiviral stock needs to be titrated. If storing for less than a day, lentivirus...For long-term storage, viral preps should be divided into single-use aliquots, and stored at -80°C. Some...want to consider concentrating your viral prep. Storage can also play a role in the success of your lentiviral...factors such as the transgene expressed, the viral storage buffers used, and other experimental parameters...dry ice/ethanol bath or liquid nitrogen prior to storing. Know your target cells Labs tend to follow the...serve as the cellular receptors for vesicular stomatitis virus.” Proceedings of the National Academy of... -
Data Freedom: The Expansion of Data Sharing in Research Publications
TypeBlog Post...publishers. Nonetheless, the initial release caused a storm as the organization left open the question of how...for advice about the scope of the policy. End of story? Not quite. The PLOS policy on data access is a...statistical methods, omission of some data, etc. The firestorm that followed implementation of the policy comprised...easily available, organized in a form that can be understood by others, balanced against how often such data...of research, raw data requires huge amounts of storage. For example, mouse behavioral geneticists may ...data should be included? We should recognize that storing data in a sharable form is indeed a significant...carried out. While no one is suggesting all data be stored in a retrievable format, we too rarely preserve... -
Why and how to start studying chromatin (with Addgene's help)
TypeBlog Post...were discussed at the 2018 Keystone meeting for Chromatin. Note that histone genes frequently have many...context of chromatin?” Though it’s true that adding histones and chromatin architecture to every diagram portraying...Chromosome Architecture and Chromosome Organization Keystone Symposium. As it happens, there are a lot of scientists...transcription of a gene a megabase away, or looking at the histone methylation status at a double-strand break that...the presentations and posters I saw at the 2018 Keystone Chromosome Architecture and Chromosome Organization...Sharon Dent Archa Fox Elaine Fuchs Kai Ge Robert Kingston Thoru Pederson Phil Sharp Julie Ahringer Alexander...RANbodies) as versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl ... -
Supporting Reproducibility with a Connected ELN
TypeBlog Post..., Word docs, and the like is likely stored in online file storage tools like Dropbox, Box, One Drive, ...commonly used online file storage tools, as illustrated in this video, and to data stored on institutional and...like image banks and sequencing files is likely stored in institutional and lab servers specifically designed... conveniently reference specific pieces of data stored externally in the record of research compiled in...fully supported API that enables development of custom integrations with other tools and resources. As... in the documents to data in the external file storage resources remain live. This means that someone ...research and commercialization community in greater Boston, while at the same time keeping one foot on the... -
Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC
TypeBlog Post...human cryostat tissue sections. J Histochem Cytochem : Off J Histochem Soc 38:923–926. https://doi.org/...Fighting with antibodies to produce immunohistochemistry images that are crisp, bright, and lacking in...avoiding background secondary binding Direct immunohistochemistry (Figure 2A) How do you avoid having your...Randolph-Habecker J (2013). A Flexible Mouse-On-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free...Immunofluorescence, and Multiple Antibody Staining. J Histochem Cytochem 62:197–204. https://doi.org/10.1369/0022155413511620...Monoclonal Antibody to Mouse Tissue Sections. J Histochem Cytochem 46:977–983. https://doi.org/10.1177/002215549804600813...Tuson JR, Pascoe EW, Jacob DA (1990) A novel immunohistochemical technique for demonstration of specific binding... -
A Quick Guide to a Career in Software Product Management
TypeBlog Post... it out. 4:00 PM--Respond to customer help tickets. Catch up on customer help tickets, and respond when...advocate for the customer and help build software and technology tools with the customer in mind. You can...Oxford Nanopore Technologies, I work with our customers in the US to learn where the gaps are in their... so many different people! I communicate with customers, software developers, as well as team members ... It’s divided into the key takeaway themes and customer pain points we’ve identified. I can’t wait to ...know what to work on next to help alleviate any customer pain points. Many thanks to our guest blogger... -
Celebrating 15 Years of Scientific Sharing
TypeBlog Post...usual quality and success. Customer and technical support Our customer and technical service brands...helpful and the customer and technical service teams put 100% effort into helping our customers every day in...have for the future. At Addgene we will never ever stop planning, perfecting, and learning, but there are...scientific fields.” Thanks to our legal team, and great customer service, somehow we have scientists actually loving...offer online for Addgenies and to serve external customers. I also can’t wait to see where our new data initiatives...to do it quickly. Hiring is hard but as a 3 year Boston Globe and BBJ Best Place to Work with a positive... -
Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute
TypeBlog Post... to learn things quickly. Story telling. The ability to know what’s a story and how to hook your readers...feels that storytelling is an impactful tool for science communicators. “The power of a story is to make...there’s always plenty of writing: plain-language stories about Fred Hutch research, pieces for donors such...that can be a very painful process. And certain stories can make you feel like ‘I’m a terrible writer. ...through connections and that’s something that good storytellers in science can do.” Susan’s number one piece...social media as a scientist here Learn more about storytelling in science ... -
Mouse Modeling, Part 1: Genetically Engineered Mice
TypeBlog Post...presence of a stop cassette upstream of the target gene in the targeting vector. The stop cassette is floxed...embryonic stem cells that are then injected into a blastocyst. There are several types of genetically engineered...Modified embryonic stem cells are injected into a blastocyst which is then implanted into a female animal ...embryonic stem cells, which are then injected into a blastocyst. Nucleases can also be used to engineer knockout...create mosaic animals where not all cells in the blastocyst were successfully edited by the injection of ... variations of this technology can be used to customize the effects of this recombination. To generate...floxed, so Cre-mediated recombination deletes the stop cassette, which permits expression of the target... -
How-to: 5 Steps to a Great Panel Discussion
TypeBlog Post...bring out these interesting stories and remind the panelists of the stories or topics you know they can... taking good notes and discovering interesting stories, riffs or opinions that each panelist can contribute...identify the funny, provocative or interesting stories that will make for a fun discussion. People are...they can see each other) on comfortable chairs (stools if you have to, but this is tough for women in ...especially short people – which I am, so I hate stools). I like it when the moderator sits in the middle...question in a way that elicits those interesting stories and opinions. Don't let all the panelists answer...moderator's job to politely but firmly get them to stop talking and redirect the conversation. The person... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog Post...production is Komagataella phaffii (a.k.a Pichia pastoris) because it secretes fewer of its own proteins...applications for validation include western blot, immunohistochemistry, and immunocytochemistry. It’s important...applications. An antibody that performs well in immunohistochemistry may not perform well in a western blot. ... to their parental hybridomas. Aggregation and storage Aggregation of antibodies may impact their quality...testing. An aliquot of our recombinant antibodies is stored at 37 °C for a period of two weeks. After this ...that are not equally effective are shipped to customers on ice, preventing any temperature shifts. Of ...we are also cautious during the aliquoting and storage process to prevent dropped tubes. Now that your... -
Transferable Skills Guide: Conflict Resolution
TypeBlog Post...primary role is customer support, but there are eight other teams who still retain a customer support element...when I discovered that many of my old suitemates stopped talking to each other if I was not there to arbitrate...to listen, and an ability to remember the right stories at the right time. Like many of the posts in our...because confirming that every person has read and understood every communication that gets sent out through...other responsibilities. Knowing which aspects of customer support each team “owns” is critical for day-to-day...-to-day operations and keeping our customers happy, but inevitably there will be gray areas that either...another job, it will be important to retain the stories and examples that have guided your resolution processes... -
Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation
TypeBlog Post...either side of a stop cassette, upstream of an effector gene. In cells where the stop cassette is removed...of cells will express the effector or retain the stop cassette following recombination. Image from Isaacman-Beck...step, the effector is expressed. In cells where the stop cassette remains, the effector is not expressed ...competing attP sites to drive expression of either a stop cassette or an effector gene. HDVR (2X Hepatitis...attP site. Truncating the attP site upstream of the stop cassette results in decreased efficiency of recombination...preferentially leads to gene constructs where the stop cassette remains following recombination. This strategy...