We narrowed to 634 results for: des.2
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TypeBlog Post...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...promotes scientific sharing The EDGE Program is designed to expand the number of systems that can be studied...genotype to phenotype studies New organismal systems described at this meeting were extraordinarily diverse. ...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...
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How to Negotiate a Successful Lab Start Up Budget
TypeBlog Post...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute...Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc. “How much do you need [for your...necessary resources. This is why at Experimental Designs Consulting (EDC), we instead have our clients focus...the institution/funding source. Major equipment includes large centrifuges, sequencers and analyzers, and... as many of these tools as possible, this even includes clocks, timers, and phones. As for reagents and... -
Plasmids 101: Broad Host Range Plasmids
TypeBlog Post...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241 ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695 Schmidhauser... meliloti, Rhodobacter sphaeroides, R. capsulatus, Rhodopseudomonas spheroides, Salmonella spp., Serratia...Brevibacterium methylicum, Caulobacter crescentus, Desulfovibrio vulgaris, Erwinia carotovora, E. chrysanthemi...meliloti, R. leguminosarum by. viciae, Rhodobacter sphaeroides, Salmonella typhimurium, Vibrio cholerae, Xanthomonas... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post... of invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized...made to destroy the invader. Wikipedia, accessed 25 November 2013. Author: James Atmos (2). Many...multiplexibility in various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas ...engineering. The diversity of the CRISPR/Cas systems provides powerful lines of defense against invading phages...CRISPR/Cas inspired genome editing was initially described by Cong et al. and Mali et al. in January 2013...Nine months later, more than 1500 publications describe work to improve the tool’s specificity, orthogonality... -
Hot Plasmids: Fall 2024
TypeBlog Post...improve solubility and stability (Figure 2). Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...promising tool for future structure-guided drug design and studies of protein ubiquitination and degradation...CRBNmidi plasmid here! Kroupova, A. et al. (2024). Design of a Cereblon construct for crystallographic and...specifically for C. elegans (Cao et al., 2024). The team designed coding sequences for mScarlet3 and mScarlet-I3... -
Antibodies 101: Reading a Flow Plot
TypeBlog Post...A-B+ cells. If you look at the dot plot in Figure 2, you might assume that all cell populations are positive... the different populations. In the case of Figure 2, one population is B- and another two are B+. Our ...are in the lower right corner. Figure 2: This flow plot has two populations that are B+ and...series of markers are set up sequentially, from broadest to most specific marker, and are represented as...are, try drawing a quadrant over the plot that divides it into four, using the location of the cell populations...Gating and plot orders As the gates progress from broadest markers to most specific, the first step is to... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog Post...under phase contrast starting day 2 after transfection. Fig: 2: Expressing mARGAna in MDA-MB-231...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the...to distinguish from background tissue unless destructive ultrasound imaging was used. We set out to develop...acoustic properties of these gas vesicles made non-destructive specific imaging in tissue difficult. The first-generation... allowed the GVs to be flexible and produce nondestructive nonlinear ultrasound contrast. It turned out... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog Post...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence... Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei...almost complete desensitization when exposed to continuous bright light. Desensitization is mediated by...discovered Marine, Anion-conducting and Intensely Desensitizing ChRs) differ from currently used ChRs in that...regulation of many genes. A number of groups have designed means of studying the effects of endogenous looping... they call the LADL Anchor. Using specifically designed gRNAs, they can then recruit these LADL Anchors... -
Visualizing Protein Turnover In Situ
TypeBlog Post...unpublished, that allowed the visualization of specific de novo synthesized proteins in situ. I was privileged..., 2010) and allows the visualization of the whole de novo synthesized proteome. However, FUNCAT alone ...PMID: 20543841. PubMed Central PMCID: PMC2920597. 2. Nathans, Daniel. "Puromycin inhibition of protein...radioactive materials is highly regulated, requires designated work areas and come with health risks. Importantly...synthesized endogenous proteins in situ. Now FUNCAT-PLA provides both quantitative and spatio-temporal information... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog Post...vector:insert ratio is used in the DpnI digestion and 2 µL of digest product are used in the transformation...Biology 38 (2016): 145-154. PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified.... Traditional restriction cloning is one of the oldest of these techniques and is a multi-step process... -
Summer SciComm: Science Storytelling
TypeBlog Post...struggle between two opposing forces or (2) the result of competing desires or (3) the presence of obstacles ... an introduction, and a resolution. Fig. 2: The structure of a story - not just for fiction! ... -
Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC
TypeBlog Post...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary. Figure 2: There are a few common solutions to avoid background...immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques,... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog Post...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi....updated Aug 20, 2020. Cas9 can be used to modify any desired genomic target provided that (1) the sequence is... may be unintentionally mutated along with the desired target locus. The PAM sequence is of particular... -
Plasmids 101: Plasmid Incompatibility
TypeBlog Post...to plasmid loss (Diaz et al., 2015). Figure 2: The partitioning problem. For low copy plasmids...of plasmid into a separate daughter cells (Figure 2) (Ebersbach et al., 2005). However, more recent studies...Plasmids. Microbiology and Molecular Biology Reviews 62(2): 434–464. Pubmed PMID: 9618448 Diaz R, Rech J, Bouet... Environmental Plasmids blog). Scientists have designed small, high copy incompatible plasmids that lead...Pubmed PMID: 25889267 Ebersbach G, Sherratt DJ, Gerdes K (2005) Partition-associated incompatibility caused... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog Post...one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth of...for C. elegans genome modification. Figure 2: Schematic from Dickinson et al. (2015) illustrating... the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C....America. To use this SEC-based system, first design and clone 500-700 bp homology arms for your gene...injected 60-80 animals per gene, they obtained the desired insertions on the first attempt for 6/7 genes. ... -
Antibodies 101: Introduction to Immunofluorescence
TypeBlog Post...counterstaining, mounting sample on slides, and confocal Figure 2: High-level overview of an indirect...into the several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, ...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment...antibody-based technique, validation and proper experimental design are essential. In this article, we’ll go through... -
A Look at Addgene's QC Process
TypeBlog Post...If a depositor provides a sequence for the plasmid, QC scientists will first align the de novo NGS result...contain the expected plasmid features. Figure 2. Addgene’s basic QC process. Created with BioRender.com...are assembled into a circularized plasmid sequence de novo (without the use of a reference sequence) using...sequence, if one was provided. If no reference is given, de novo assembly in Geneious can still provide additional...plasmids in our collection. This standardization provides security for requestors, ensuring that the plasmid...types of data for each of their plasmids, from descriptions of key components of the plasmids to partial...believe may affect the function of the plasmid. This includes point mutations, frame shifts, missing elements... -
Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science
TypeBlog Post... go back and finish the review. Peer-review step 2: Detailed assessment After this initial assessment...salt-induced changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced sodium...of my review - "The manuscript by ZZZ et al., describes ... The authors share a wealth of data on ... ...Additionally, you can ask that data and resources described in the manuscript are accessible to the community... -
New and Upcoming Viral Vectors - June 2019
TypeBlog Post... AAV9, AAVrg pAAV-hSyn-DIO-mCherry 37825 AAV1, 2, 5, 8, 9 (20 µL size serotype testing) pAAV-CAG-GFP...size serotype testing) pAAV-CAG-GFP 50465 AAV1, 2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP...best serotype for their particular study, which includes comparing the transduction patterns of different...fluorescence of the sensor. This “contrast” variant provides improved detection for tracking large populations... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor... determine if proteins are targeted to their destination co- or post-translationally. TRICK: Visualizing... usually translated in a specific compartment. Despite these limitations, these techniques represent powerful...