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...offerings! We recently re-amplified our distribution stock of the Brunello Human gRNA library, and we thought... a file in FASTQ format, a standard format for storing NGS reads and their corresponding sequencing quality...in an automated manner. Addgene commonly uses a custom script derived from the count_spacers.py file from...representation of your library. When plotted as a histogram, the counts will give you a bell curve of the ...finds that this representation is most easily understood visually. As an example, the curves from our ...screening, and if you have any tips, input, or stories of interest about library use, let us know in the...

...help win a business plan competition and advance investors to the next stage of diligence." In business competitions...non-scientific audiences. "There are 100 ways to tell a story - convince your audience you are doing amazing things...The team is key, and the initial team is the cornerstone of it all. Without a good nucleus, the company...are 100X better than current technology and get customers to change behavior. When you are ready to commit...crowd funding, federal funding, foundations, and investors are examples of various sources of funding. Always... Biology & the Importance of Assembly Standards Story of a SynBio Startup: RevBio's Epiphany (or Lack ...

...privilege to find and share what stories we can, and to hear the stories others have to tell.  "Also thanks...we wanted to walk through some of our favorite milestones and share some thoughts from the community that...six years to reach our first major repository milestone — 10,000 plasmids deposited and available — and... which was a major barrier for depositors and requestors alike. We transformed this process by aligning...Addgene reached the 500,000 plasmids distributed milestone! We were thrilled to see the impact our repository...

... Outlines a detailed scope of specific aims, milestones, possible setbacks, and potential alternative...Carefully read over all parts until all aspects are understood. While reading the funding description, pay attention...will address these limitations and lead to new milestones in the field. Research strategy The Research...aims The Specific Aims section highlights each milestone that must be met in order to deem your proposal...proposals consist of 3 to 5 aims, with secondary milestones to meet along the way to achieving each aim. ...comments may make it clear that the reviewer misunderstood the point that was being made, take it as an...

.... The viral genome DNA is further protected by histone-like proteins forming the nucleocapsid. B) Three...recommended to test for the presence of RCA in the viral stock by doing a plaque formation assay on A549 cells ...recombination for amplification. The final purified plasmid stock should be analyzed by endonuclease restriction digest...you’ll produce the primary recombinant adenoviral stock (rAdV-S). HEK293 cells are transfected with the ...lysate is your primary low titer rAdV-S, and can be stored at -80C for later use or used immediately for amplification.... Purified high-titer rAdV is then dialyzed and stored at -80C. Purification and concentration kits that...

...advocating-for-science-boston-2016/. Sign Up For the Advocating for Science Symposium Boston 2016 Changes since...Science Symposium and Workshop 2016 – Sept 16-17 MIT, Boston The Advocating for Science Symposium and Workshop...also started progress towards solutions. The 2015 Boston FoR meeting was designed to explore areas where...various levels, with panelists including: Kate Stoll, PhD - Senior Policy Advisor MIT Washington Office...Professor of Sociology. University of Massachusetts Boston Christine Oslowski, PhD - Biotechnology Communications...

...cause bad reactions in mice. Customization AAV offers a level of customization that is unmatched by other...over the past decade. CRISPR screens are easily customizable and enable the targeting of hundreds to tens...lentivirus but offer an improved safety profile, customization, and enhanced expression in many mouse tissues...combination of serotypes and psuedotypes means you can customize your CRISPR screen to a high degree. This is particularly...interrogate immune cells to improve immunotherapy in glioblastoma, and generate programmable screening platforms...

...Harris Lab relies on the restoration of a frame-shift in mCherry that restores fluorescence to monitor ...that disrupts fluorescence due to a frameshift. Restoration of fluorescence can only occur through editing...results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using...C-to-T editing by the APOBEC-Cas9 editosome would restore eGFP fluorescence, making it easy to quantify single... point mutations in eGFP that results in the restoration of fluorescence. Transient reporter for editing...

...proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old antibody...on-target delivery of genetic cargo  By Alyssa NeuhausTo improve delivery and reduce off-target effects....1101/2023.02.07.527548.    Tracking cellular history with protein ticker tape By Brian O’Neill The Cohen...created a system for recording the transcriptional history of a cell. Lin et al. generated a fusion protein...ticker tape images could recover transcriptional histories, accurate to within 40 minutes. Figure ...their initial tests with thick slice (70 μm) immunohistochemistry (IHC) (Figure 5). PSD-95 is a scaffolding...

...control by Addgene before being distributed to customers. Details about our production protocols, titering...vivo studies. Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by...preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins VP1, VP2...The preparations are then aliquoted, frozen, and stored at -80 °C. Concentrated lentiviral preparations...then resuspended in PBS, aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral... lentiviral preparations that have been stored at -80 °C and thawed. This ensures that any loss of titer...

...structures (at < 5 Å resolution) of the linker histone H1.8 in Xenopus nucleosomes and in complex with...has developed a new epigenetic editor, Coupled Histone tail for Autoinhibition Release of Methyltransferase...can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate the endogenous methyltransferase...uses a fusion of the DNMT3L cofactor (D3L) with a histone tail (H3K4me0) to a DNA-binding protein (dCas9 ...team to develop self-silencing CHARM variants that stop their own expression after silencing their target... a familiar challenge for anyone who has done histology on brain tissue is identifying the cells you are...

.... We are the Customer Support Specialists here at Addgene. If you’ve called our customer service line ...'Regan Ordering jeans online from your favorite store is easy. Ordering genes on plasmids online is a ...really getting at one bigger question: How does a customer place an Addgene order using a PO number? Why ...full of information about how to deposit plasmids, store your plasmids or viral items, technical support,...

... Center for Regenerative Medicine of Boston University and Boston Medical Center. Special thanks to our...director of the Center for Regenerative Medicine at Boston University Medical School. Darrell’s research focuses...Chapter 1: Introduction to Darrell Kotton and the Boston University Center for Regenerative Medicine. Darrell...Center, and the Pulmonary outpatient clinic, all at Boston Medical Center. In addition he conducts basic stem...

...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and...ligated into the MCS/SacI site in pUCXKT, removing the stop codons during backbone digestion and providing the...Toolkit for constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab...Technologies. The MultiSite Gateway Kit is a customizable Gateway®-based toolkit that allows scientists...bound to the mRNA since they are tethered AFTER the stop codon. Thus a non-translated mRNA will appear yellow...: 10.1126/science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector...LacZ, puromycin or CreERT2 can be combined with custom-cloned vectors in a Multisite Gateway reaction ...

...my training, I dove straight into my capstone project. A capstone project is a major project given to ...Figure 1: Addgene's summer interns presenting their capstone projects during the Summer Intern Poster Presentation... scientist. But I expected something wrong. My capstone project was based on plasmid cloning and I, the... Once everything was confirmed, I worked on my capstone project report and presentation, during the final...

...you questions. Remember that people like a good story and science involves characters – in this case you...or other places online, and consider how those stories are being told. Twitter can be a good way to follow...surprised to find that his friends and family finally understood why he cared to study mice on some distant island...the East Coast, there’s programs at both MIT and Boston University. At Addgene we're committed to scientific...sharing – both your plasmids and your research stories. Consider practicing communicating your research...

...Emission Depletion (STED), and Stochastic Optical Reconstruction Microscopy (STORM). These dyes have specific...Throughout their history, fluorescent dyes have enabled the visualization of both organic tissues and...than was absorbed. This phenomenon is known as the Stokes Shift, or the difference in wavelength between ...technologies in fluorescent dyes/ligands Throughout the history of cell biology, fluorescent labeling techniques...fluorescent protein Fluorescent Proteins 101: A History of Fluorescent Proteins ...

...IP and ChIP assays. Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP...first step of optimization, we started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments...tagging, pick a crRNA that cuts 8-15 bp downstream of Stop codon; for N-terminus tagging, a crRNA cutting 8...csRNP experiments, it can be ordered in bulk and stored in small aliquots at -80 oC. Assemble csRNP: Anneal...near the insertion site (within ~8-15 bp from the stop codon for C-terminus tagging) GC content of the ... hands, delivering Cas9 protein in complex with custom synthetic cr:tracrRNAs (csRNP) is the most efficient...

...not necessarily represent the views of Addgene. Boston, MA, USA 42.3601°N 71.0589°W – Alina Chan The ...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...to join the public conversation and to tell the stories behind our research.  That's why I marched for ...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...behind these two fingers the ninety-nine cubits of Aristotle, nor would you mention my small error and at the...second and that it is our duty to tell our science stories in a manner that encourages open discourse with...

...skeptical reader of historical fiction (I can personally attest to this!). Following history from the Ice Age...Wrong - and the New Research That's Rewriting the Story by Angela Saini "This book is a smart and carefully...Strange & Mr. Norrell by Susanna Clarke "A quasi-historical/fantasy fiction. Two dueling magicians in the...of England by Edward Rutherfurd "A 10,000 year history of southern England. While the length may be intimidating...