We narrowed to 492 results for: ara-2
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TypeBlog Post...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...science.aav4294 Ozcan, Ahsen, Krajeski, Rohan, Ioannidi, Eleonara, et al., Programmable RNA targeting with the single-protein...
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Troubleshooting and Optimizing a Western Blot
TypeBlog Post... that all look exactly alike. Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...buffer. You’ll want to run long enough to get good separation in your ladder, particularly in the sizes around... -
With an Eye Towards the Future, We Look Back at the March for Science
TypeBlog Post...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...not only believe in the separation of church and state, but also the separation of fact and fiction. Our... -
Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins
TypeBlog Post... fluorescent proteins expressed in the specimen. 2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil... tagged with a fluorescent protein or dye. It separates the cells mechanically in a vibrating nozzle, ...Schaeffer, Marie, et al. "Dynamic imaging of T cell-parasite interactions in the brains of mice chronically... -
Optogenetics Guide
TypeGuide...Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein interactions based on Arabidopsis thaliana... OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated forms of...tools derived from direct comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551...influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool,...thaliana cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly sequestering targets into...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control... with single-spike temporal resolution. PLoS One. 2(3):e299. PMID 17375185 Kim B, Lin MZ. 2013. Optobiology... -
Adenovirus Guide
TypeGuide...use of pAdEasy-2 (Addgene #16401) can increase the capacity of the rAdV vector. pAdEasy-2 does not contain... incorporate additional genomic deletions. Figure 2: First-generation rAdV vectors. Created with BioRender.com...and have to be handled at biosafety level two (BSL-2). They are highly immunogenic and can trigger a strong...era of personalized medicine . Genes & Diseases, 4 (2), 43–63. https://doi.org/10.1016/j.gendis.2017.04.001...adenoviruses using the AdEasy system . Nature Protocols, 2 (5), 1236–1247. https://doi.org/10.1038/nprot.2007.135...Adenoviral vector vaccine platforms in the SARS-CoV-2 pandemic . NPJ Vaccines, 6 (1), 97. https://doi.org...Adenoviral backbone plasmid that lacks E1 and E3. pAdEasy-2 Adenoviral backbone plasmid that lacks E1, E3, and... -
Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart
TypeBlog Post...Joachim ended up at the University of Amsterdam Chapter 2 - Joachim’s Academic and career goals. Career advice...microscopy (University of Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes... -
Modular Cloning Guide
TypeGuide...assembled into a Level 2 vector, forming a functional genetic circuit. Level 2 vectors are often designed...of three sets of cloning vectors (Level 0, 1, and 2) which can be used in successive assembly steps. Before... iterations of assembly. Combining multiple Level 2 vectors permits the creation of even more complex ... 16 RBS strength variants, 8 tag-compatible RBSs, 2 secretion tags, 6 CDSs, 9 terminators, 4 nonfunctional...of multigene constructs. PLoS One . 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PMID: ... Expression John Dueber 96 standardized and characterized parts that can be used for bottom-up hierarchical...constructs expressing GFP at 18 levels for rapid characterization in new organisms. Mobius Assembly Vector Toolkit... -
QC Sequencing Technologies at Addgene
TypeBlog Post...short-read sequencing are highlighted in Figure 2. Figure 2: Overview of short-read sequencing technology...synthesized (Figure 1). Sanger sequencing requires four separate reactions, where a fraction of one of the four...The resulting DNA fragments are pooled and then separated by size using electrophoresis, typically capillary...deconvoluted to identify the bases (and, if necessary, separate different samples), the resulting data, or reads...nucleotide that passed through based on these characteristic drops. Figure 3: Overview of Nanopore...To address this issue, the QC team developed a separate script for manually inputting known innocuous ... -
Antibodies 101: Immunoprecipitation
TypeBlog Post...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...controls for an IP. When possible, perform the IP in parallel with both a sample known to express the protein...identify any non-specific interactions, perform parallel IPs with beads only and an isotype control antibody... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because ‘antibiotic...Resistance can be intrinsic, meaning a basic characteristic that naturally occurs in the bacterium, and... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post...from animals or hybridomas. Figure 2: (Left) Figure 2: (Left) The light chain and heavy chain genes..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P... -
Plasmids 101: Terminators and PolyA signals
TypeBlog Post...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...and translation processes as well. Figure 2: The conserved eukaryotic polyadenylation signal directs...transcriptional read-through. Chris Voigt’s lab has characterized a set of prokaryotic terminators and deposited... -
Antibodies 101: Conventional vs Spectral Flow Cytometry
TypeBlog Post...from another within a fully stained sample. Figure 2 is an example of the emission spectra for two fluorophores...another during spectral unmixing. Figure 2: The full emission spectra for BV421 and BV711 on ...experiment, it is important to consider how many parameters you want to use in a given experiment. For very... -
Antibodies 101: Antibody Engineering and Directed Evolution
TypeBlog Post... follow the same general cyclical process (Figure 2). The differences typically come in when generating...starting point for the next cycle. Figure 2: General process of antibody directed evolution. Created...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...antibodies developed by IPI are extensively characterized and validated, with the application data from... -
Plasmids 101: Stringent Regulation of Replication
TypeBlog Post...the chromosome, ending replication. Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...cooperatively to separate nearby AT-rich regions of DNA. In a test tube, the DNA helix can be separated by DnaA...ATP-bound DnaA, the proteins cooperatively can separate the DNA strands. Created with BioRender.com. ... -
CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!
TypeBlog Post...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...once), however, SAM exhibits activation levels comparable to other popular activation methods (VPR and ... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog Post... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...10634178. 8. Murnane, John P., Michael J. Yezzi, and Barbara R. Young. "Recombination events during integration... -
PCR Applications for Copy Number Variation Assays
TypeBlog Post...mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so forth....quantitative PCR. Expert Rev Mol Diagn. 2005 Mar;5(2):209-19. doi: 10.1586/14737159.5.2.209. PMID: 15833050...falciparum multidrug resistance-1 gene (pfmdr1). J Parasitol. 2011 Oct;97(5):939-42. doi: 10.1645/GE-2792.1... -
"Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science
TypeBlog Post...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...stereotaxically, STX) to the brain. Figure 2: Selection, prep, and testing of AAV enhancers in ...B., Ben-Simon, Y., Opitz-Araya, X., Martinez, R. A., Way, S. W., Thyagarajan, B., Otto, S., … Ting, J....Resources References Ben-Simon, Y., Hooper, M., Narayan, S., Daigle, T. L., Dwivedi, D., Way, S. W., Oster... J. K., Taormina, M. J., Martinez, R. A., Opitz-Araya, X., Roth, J. R., Alexander, J. R., Allen, S., Amster...E. E., Mahoney, J. T., Kojima, Y., Ding, Y., Somasundaram, S., Miller, J. A., Kalmbach, B. E., Radaelli...