We narrowed to 833 results for: ins
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TypeBlog PostPublishedMay 24, 2018, 1:20 p.m.... they bind to a class of Argonaute proteins. The Argonaute proteins remove one strand of the siRNA and...cDNA), and then use PCR and molecular cloning to insert the cDNA into the desired plasmid backbone. The...the process is a little more complicated. These strains are more picky eaters and refuse to eat liver paste...Sánchez lab protocol also provides step-by-step instructions for the “the bravest and more curious” students...
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5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m....use a standard strain, like DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity ...prefer to store freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting...option may be to avoid freezing altogether, and instead to use the virus immediately upon harvesting. Interestingly...appear as a white pellet. The pellet can then be rinsed and resuspended in cold sterile PBS. Alternatively...of ensuring viral production is to perform PCR against components of the viral vector using the harvested... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog PostPublishedDec. 1, 2015, 3:30 p.m....Director of the National Human Genome Research Institute (NHGRI) of the NIH, started off with the first...will lead to advances in human health. It was inspiring to hear how the genome project has already had...enzymatic machinery (i.e. RNA replication with no proteins). The chemistry is complex and this work requires...desribed how P. furiosus has systems to defend against both DNA and RNA viruses - 2 cas systems regulate...system. His lab is also exploring the use of cas9 proteins from different species for genome engineering.... -
The Materials Science of Optogenetics Experiments
TypeBlog PostPublishedSept. 17, 2015, 2:30 p.m....—an optical insulator called a ferrule (below). The fiber optic cable/ferrule is inserted into the target...stimulation guarantees that the majority of the opsins will receive sufficient quantities of photons to...although you may be able to purchase one pre-made instead). Laser stimulation In the video above...: closed/light receptive, open, closed/light insensitive. If you deliver a continuous stream of photons... -
The Scientific Conference Poster Session: Tips for Success
TypeBlog PostPublishedAug. 29, 2019, 12:50 p.m....practice: Rehearse your presentation with people inside and outside your field. Your colleagues, lab mates... tack up your poster seconds before the session begins. Plus, you’ll have extra time to practice at the... in to the next figure. Remember your outline: Instead of getting bogged down in the minutia or coming... practicing with your colleagues, ask them to brainstorm potential questions you may receive. People often...in to thirty-minute-long presentations, which constrains the number of posters you can see. If you are... -
Pooled Library Amplifications
TypeBlog PostPublishedMay 30, 2024, 1:30 p.m....variance of size of plasmids in your library (Are the inserts of different sizes?), and the desired coverage ...100x coverage of your library, and your library contains 5,000 plasmids, your goal for your transformation...replicate faster. For this reason, if your library has inserts of different sizes (such as a transcription factor...library is of a smaller size with a single size insert, liquid media can be faster and easier to work ...protocol paper by the Zhang lab, and the Broad Institute’s general pDNA Library Amplification protocol are... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog PostPublishedJuly 30, 2020, 1:15 p.m....previous books focused primarily on fluorescent proteins and their applications, tackles much broader issues...from mildly surprising to genuinely shocking. Mainstream media outlets have addressed CRISPR frequently...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...Addgenies Joanne Kamens, Marcy Patrick, Aliyah Weinstein, Fabienne Furt, and Quintin Marcelino for their... Eric J. Perkins... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m....pastoris) because it secretes fewer of its own proteins, making the purification process simpler. Due ... the production of Fab fragments where variable chains are secreted into the periplasmic space to be oxidized...SDS-PAGE gel. Once stained, the heavy and light chains become visible alongside your ladder and a protein... density of the antibody bands can be measured against the protein standard and the concentration may ...Any bands on the gel beside the heavy and light chains can also be measured. Contaminating bands lower... -
Uncovering Ancestral Bias in CRISPR Research
TypeBlog PostPublishedNov. 29, 2022, 2:15 p.m.... for instance, a recent preprint released by the Boehm and Beroukhim labs at the Broad Institute of MIT...MIT and Harvard, Dana-Farber Cancer Institute, and Koch Institute, where an exploration into the effects...introduced an ancestry bias into their experiments instead of a true correlation. It turned out there was... -
MXS Chaining
TypeBlog PostPublishedFeb. 7, 2017, 3:30 p.m....sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies...modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals...combined to create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa...later combine all four constructs into one 15 kb insert. Table 1: MXS Constructs for Labeling Cellular ... fluorescent proteins... -
CRISPR 101: Any Base Transversion Editors
TypeBlog PostPublishedApril 1, 2025, 1:15 p.m....DNA N-glycosylase (UNG) enzymes. Various other proteins included in different CGBE tools allow the researcher...a similar tool shortly thereafter, using mouse instead of human AAG (remember, MPG and AAG are the same...a more specific A → C base editor (ACBE), they inserted the deaminase into the C-terminal domain of Cas9...excises inosine can be modified to target guanosine instead, perhaps analogous engineering of UNG, which excises...its homologs) has more than one name. Indel Insertion/deletion A type of unwanted DNA edit. AYBE, ... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog PostPublishedJan. 24, 2023, 2:15 p.m....cassette contains the major structural gene gvpA marked with EBFP2 and the second cassette contains the P2A-linked... be easily expressed heterologously by bacteria inside the body. So we screened gas vesicles gene clusters...bARGSer-expressing E. coli. The GV-specific ultrasound signal inside the tumors was overlaid with the anatomical B-mode...which seemed overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog PostPublishedMay 8, 2018, 1 p.m.... thanks to our guest blogger, Jessica Roginsky. Jessica Roginsky leads the scientific support team at ...This article was contributed by Jessica Roginsky, Scientific Support Lead at Synthego. Article source...genomes that have been successfully modified with insertions or deletions (indels) and then characterizes ...each bar of the indel plot shows the size of the insertion or deletion along with the percentage of genomes... -
Transferable Skills Guide: External Collaborations
TypeBlog PostPublishedApril 19, 2018, 1:25 p.m....collaborations. During my graduate studies at Karolinska Institutet I started working with other groups. First...minutes. Get to know your collaborators from other institutions personally on conference calls and make sure...learning and, in my experience, mistakes and insecurities are more easily excused. When you reach out ...Collaborate: An Imperative for Graduate Students" from Inside Higher Ed Additional Resources on the Addgene Blog... -
How to Write a Scientific Review Article
TypeBlog PostPublishedFeb. 16, 2017, 3:30 p.m.... of all the proteins in the class that showed the relative sizes and the conserved domains. This information...highlighted a few dozen things that I didn’t know about. Instead of reading a paper and getting my bearings, I would...summarized how frequently each member of a class of proteins was mutated, as reported in various studies. We...sense of the similarities and differences among proteins in the class. Offer your perspective It doesn... -
Using Video to Share Your Science: We Share Science
TypeBlog PostPublishedNov. 3, 2016, 2:30 p.m.... This post was contributed by Ryan Watkins, Professor at George Washington University and developer of...in scientific journals flies under the radar and gains little notice. A 2009 research study found that ...influenced by research in other disciplines. But there remains a great deal to learn from the science of others...platforms (such as social networking, visual appeal, instant sharing across multiple other sites), I created...and let the world know about your science! Ryan Watkins is a professor at George Washington University ... -
Tips for CRISPR Gene Editing in Mice
TypeBlog PostPublishedJune 28, 2016, 10:59 a.m....Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains overlapping 5′ and 3′ EGFP fragments under the...MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and sgRNA expression... surrounding your gRNA. Once amplified, you can insert this region into a pCAG-EGXXFP plasmid using standard...techniques (The XX indicates where your target is inserted, disrupting the EGFP fluorescent signal, for details...give specific details on what exactly has been inserted or deleted. Each lab will use a slightly different... -
The Challenges of Cell Culture
TypeBlog PostPublishedMay 5, 2016, 2:30 p.m....Science (DBCLS), National Cancer Institute, Public Domain, National Institutes of Health, Wikipedia user Aimecie...isoenzymes and genetic polymorphisms specific to their origins. This technique is still used today to test for...working cell bank has been made by a research institution (an excellent practice), or maybe cells have ...Brendan P., Walter A. Nelson-Rees, and Grover M. Hutchins. "Henrietta Lacks, HeLa cells, and cell culture... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m.... the study of neurological diseases, such as Parkinson’s disease (PD). PD is primarily characterized by...scientists to easily observe fluorescently labelled proteins and tissues to better assess developmental processes...of the pancreas and produce different levels of insulin (Hesselson et al., 2009). The worm (Caenorhabditis...From the baker to the bedside: yeast models of Parkinson's disease." Microbial cell 2.8 (2015): 262. PubMed...quick guide to Drosophila Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources... -
Transferable Skills Guide: Conflict Resolution
TypeBlog PostPublishedJan. 4, 2022, 2:15 p.m....or projects is a common source of conflict. For instance, at Addgene, there are two teams whose primary... will not be great at resolving conflicts. Institutional Knowledge: Of the tools I’m mentioning here,...accomplish, but there is a potential for long-term gains when you can do it. If Party X proposes solution...have used your empathy, listening skills, and institutional knowledge to resolve conflicts. Conflict in ... Eric J. Perkins...