We narrowed to 522 results for: des.2
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TypeBlog PostPublishedMarch 30, 2023, 1:15 p.m....responsible for that differentiation. Fig. 2: Single cell RNA-seq clustering of cells transduced...transcription factor library The human genome encodes approximately 1,800 TFs, with >3,500 unique isoforms... and screening mechanisms, although the unique barcodes for each TF help streamline identification. While...a disease in a model system. TF screens can be designed to identify custom TF programs to acutely initiate...
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Communicating Your Science With Help From ComSciCon
TypeBlog PostPublishedJuly 19, 2018, 1:12 p.m....aspects of the work you presented may also be unclear. 2. Make the audience your primary focus When writing... craft sentences for abstracts and presentation slides, learn from audiences as they do science outreach...about whether the same principle applies to other slides in your deck. If someone asks a question about...of a new science communication initiative that provides yet more scientists with opportunities to practice... -
Antibody Validation for Flow Cytometry
TypeBlog PostPublishedSept. 10, 2024, 1:15 p.m....types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube. Figure 2: Schematic showing the use of an orthogonal approach...see: Uhlen et al., 2016). In this post, we will describe the approaches that can be used to determine selectivity...intended application — here, flow cytometry — is the desired target, using one or more of the approaches outlined...interpreting your data therefore necessitates careful design of RNAi sequences, confirmation of RNA knockdown...the target at different levels, it is possible to design experiments where the performance of the antibody...We find that cell tracker dyes are helpful in designing such panels, so that pre-stained cell lines can... -
Plasmids 101: Biotinylation
TypeBlog PostPublishedNov. 15, 2018, 1:50 p.m....Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...26224555. PubMed Central PMCID: PMC4884002. 10. Boer, E. de, Rodriguez, P., Bonte, E., Krijgsveld, J., Katsantoni...Northern blotting Cell surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated...your downstream application. Biotin labelled oligonucleotides can be used in a multitude of applications...commonly used this method to attach proteins or polypeptides to a solid substrate (Holden and Cremer, 2003... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog PostPublishedNov. 6, 2015, 3:30 p.m....University of Oviedo, the European University Miguel de Cervantes, the University of Navarra, and the University...15.4%) 46 (59%) of these students were in the last 2 years of their degree. Students filled in a pre-program...together with the universities, a bright future. IMP provides matched one-on-one international mentorship, with...recommended, a mentor with international experience provides his/her mentees with a broader, first-hand vision... -
K. phaffii: Rising to the Occasion
TypeBlog PostPublishedApril 4, 2023, 1 p.m....work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...biomass production techniques in the following decades, Phillips made the decision to release the K. phaffii...of K. phaffii lacks certain terminal mannose glycopeptides (which most other yeast possess) that are problematic... -
Plasmids 101: Mammalian Vectors
TypeBlog PostPublishedMarch 25, 2014, 3:15 p.m.... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK...mammalian plasmid components, it may be useful to describe the means of introducing genetic material (such...the plasmid in the same way. Selection methods, described below, are typically employed when creating stable... -
AAV Q&A with Tim Miles
TypeBlog PostPublishedMay 17, 2022, 1:15 p.m....retransduce a model organism such as mice, e.g. first at 2 months, then at 4 months? Or do antibodies become ...1x1012 vg/mouse is more standard) and reflects a desire for very high MOI on top of very high coverage....researchers have used the flexibility of AAV genome design in remarkably creative ways. The continued development... -
Hot Plasmids - May 2022
TypeBlog PostPublishedMay 26, 2022, 1:15 p.m.... those that also infect the liver. Fig. 2: Summary of the generational evolution of the AAV ... error rate for long sequence knock-ins Visual barcodes for live cell clonal multiplexing Improving ...nanobody-GFP interaction). The modularity of the system provides researchers better flexibility and makes the approach...other experiments. And, while the BLITZ system was designed and optimized with zebrafish in mind, it can easily...https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal multiplexing by: Rachel Leeson...Recently, Ravid Straussman’s lab has developed visual barcodes for clonal multiplexing of cells in live imaging...peptide. By combining three different localization peptides with four different fluorescent proteins, and ... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m.... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While two... two guides require two arrays for Cas9, two guides can be streamlined into a single array for Cas12a....applications. This set of new anti-FLAG frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag ...target multiple genes within a single cassette provides increased simplicity for combinatorial genetic...cost-effective alternative to commercial ladders which also provides a useful yet low-pressure teaching opportunity...less than half the size of either Cas9 or Cas12a. Despite its small size, CasMINI is comparable in efficacy... -
Addgene's Tips for Plasmid Quality Control
TypeBlog PostPublishedJan. 14, 2016, 3:30 p.m.... and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above...sometimes have to design new primers to sequence the important features. You can use primer design software such...such as Primer3 to help you design your custom primers. Remember, when designing a sequencing primer, you... to differ between plasmids, and selecting or designing a primer specifically to target this region, as...instructions on other factors to consider when designing your primers can be found here. Finally, you will... to Verify Your Plasmid Learn How Addgene Uses Barcodes to Track Samples Resources on the Addgene Website... -
Plasmids 101: Screening Strategies Used in Plasmid Cloning
TypeBlog PostPublishedSept. 15, 2020, 1:15 p.m....about colony PCR in this blog post. Figure 2: The steps of colony PCR. Sanger sequencing Sanger...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a screen comes in. Let’s...blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse...Sanger sequencing determines the precise order of nucleotides within a DNA molecule, in this case a plasmid... of the information you need, so you'll want to design a custom primer. Because Sanger sequencing can ... -
All in a Twist: dsRNA
TypeBlog PostPublishedFeb. 2, 2023, 2:15 p.m.... would release them into the cytosol. Fig. 2: Endogenous sources of duplexed RNA DNA:RNA hybrids...into agriculture as species-specific pesticides and insecticides, an advancement over previous chemical...prone to degradation – those are the words that describe RNA! Double stranded? Well, that’s just for DNA...also known as RNAi, are short, dsRNA molecules designed to degrade mRNA transcripts. RNAi activates the...sequence is found (typically RNA that the RNAi is designed against), RISC can repress translation and/or ... and repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular... -
Gendered Innovations: Why Does Sex of the Cell Matter?
TypeBlog PostPublishedJuly 21, 2016, 2:30 p.m....state-of-the-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on increasing...using both sexes in basic research is excellent design), but they used all female stem cells—this was ...environmental factors. How to include sex in experimental design Recognizing the importance of analyzing sex, the... above, another excellent resource for research design is Kalpit Shah et al., “Do you know the sex of ...Corporations need to integrate these insights into product design. But eyes have been opened—and we cannot return... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog PostPublishedFeb. 28, 2023, 2:15 p.m.... high-priced commercial devices. Fig. 2: Workflow for open software (left) vs. open hardware...provide the construction plans or CAD (computer aided design) drawings of their devices, which makes it very...has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally established...similar way to the biological protocol, and assembly designs based around the use of commercially available ...users outside the project to modify the raw and design files, allowing the device to take on a community-driven...of work from universities and research centers provides a potential conflict between traditional IP-based... -
Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity
TypeBlog PostPublishedNov. 5, 2019, 1:28 p.m....divided by the off-target activity. Figure 2: Specificity is the measure of off-target activity...Requirement and Expanding CRISPR Beyond SpCas9 described the PAM specificity of Cas9s isolated from different..., and NGCG). Since then, the Osamu Nureki Lab described a rationally engineered Cas9, SpCas9-NG, which...Nishimasu et al., 2018). The David Liu Lab similarly described xCas9, which displayed the ability to target multiple... also high off-target activity, which may be undesirable in many applications. At the time of our first...post on choosing a Cas9, the Feng Zhang Lab had described eSpCas9, while the Keith Joung Lab had developed... -
Plasmids 101: Yeast Vectors
TypeBlog PostPublishedFeb. 25, 2014, 7:11 p.m....and are considered “high copy”. A fragment from the 2 micron circle (a natural yeast plasmid) allows for...element. This link provides a more extensive list of yeast auxotrophic markers and includes the associated...differ from the bacterial elements previously described. Why do scientists use yeast vectors? Yeast are... that come with working with prokaryotes. This includes but is not limited to: rapid growth, ease of replica...most commonly used selection markers in yeast and provides the element needed to overcome the auxotrophy ... -
Summer SciComm: Preprints
TypeBlog PostPublishedAug. 16, 2022, 1:15 p.m.... scientific communication ecosystem Figure 2: The scientific ecosystem, enriched by open access...been common in fields like math and physics for decades (see: arXiv.org), but have been only adopted by...course, preprints don’t replace journals and other modes of scientific communication. Researchers often post...to receive deposits of plasmids that have been described in a preprint. In fact, over 1,500 plasmids in...Depositing plasmids that were used in a preprint provides all the same benefits as sharing those plasmids...resources on the Addgene blog Summer SciComm Series: Modes of Communication Intro to Our Summer SciComm Series... -
Using CRISPR/Cas9 to Edit Disease Out of the Genome
TypeBlog PostPublishedJune 23, 2022, 1:15 p.m....cellular physiology. https://doi.org/10.1002/jcp.30064 2. Barman et al. 2020. “A glance at genome editing with...ability to influence editing efficiency. In 2022, despite the increasing effectiveness of CRISPR/Cas9, scientists... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog PostPublishedMarch 14, 2019, 1:54 p.m.... enrich the microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol...producer cells and purification protocols often describe a process that begins with cell lysis to release...