...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...

...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...

...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...

...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...

...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...

... 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...

...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 ...

..., 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...

...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...

... 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...

... 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 ...

...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...

...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...

...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...

...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...

...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...

...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...

...would love to tell such a dramatic story, but it wouldn’t be my story. I love academics. If I didn’t, it...there was even a time when I was too nervous to ask store clerks for help at the mall. In undergrad I was ...to open up a little after I dropped myself into Boston University with none of my old friends to fall ...thought that I would be best poised to share the stories and difficulties behind great discoveries by tackling...actually finishing up this post on a plane back to Boston from Kentucky where I spent a couple of days talking... mission to make research easier. I’m hoping my story demonstrates that transitioning from academia doesn...

...fresh stocks. Most antibiotics are stable in powder form, but quickly breakdown in solution. Storing aliquots...them to media hotter than about 55oC and store plates/stocks wrapped in foil if a light-sensitive antibiotic...tool in molecular biology, yet scientists rarely stop to think about how much easier it makes our lives.... For instructions on how to prepare antibiotic stocks, see Addgene's Reference Page. How else can antibiotics...antibiotics be used in the lab? Historically, antibiotics have also been used to disrupt genes at the chromosomal...

...their CPP system to study protein transduction, D’Astolfo et al. made the surprising discovery that both...NDSB-201) were required for protein delivery. D’Astolfo et al. coined the term iTOP (induced transduction...that iTOP functions through macropinocytosis. D’Astolfo et al. tested iTOP in a series of primary cell...resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP...Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to see if iTOP would be compatible...cleave the specified target. Figure adapted from D’Astolfo et al. In certain instances, the potential...Efficient Intracellular Delivery of Native Proteins. D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR, Clevers ...