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Showing: 271 - 300 of 844 results
  1. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    ...cartoon of the protein.) In the middle of my presentation, lost in a sea of data, my mind went completely...taught me an incredibly valuable lesson: Your presentation will be better if you take time when you need...need it, rather than trying to push through a state of panic. Now, if I feel myself getting too nervous...sense. As tempting as option two was in my panicked state, only option one would let me accomplish my goal...often as needed until you get to the end of the presentation, at which point you will have achieved your ...gig. Being able to give a clear, informative presentation, speak at conferences, or even just provide ...
  2. What's New in CRISPR - November 2021

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    Blog Post
    ...dominant negative mutation in MLH1 (PE4 = PE2+MLH1dn and PE5 = PE3+MLH1dn). Silent mutations added near the...the Cas12f scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI...  and base editing, whereas a different set of mutations (the “V3.1” version) works best for genome editing...protein PEG10 specifically binds its own mRNA to facilitate its packaging into virus-like particles. Feng...boosts the efficiency of prime editing to correct mutations in a variety of target genes and cell types. Their...
  3. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

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    Blog Post
    ...articles. Then just start reading and following citations through the literature until you have all of the...they mean. Could there be any other possible interpretation of the results? Then think deeper about the...Is the data of sufficient quality (error bars, statistics, clarity in images etc)? Did they perform all...doesn’t mean that there are no mistakes or misinterpretations. After all, the science was performed, written... brim with reprints and papers with scribbled notations in the margins. If you prefer, printing and hand-writing... Department of Biological Sciences at Louisiana State University. She is a fan of photosynthesis and writes...
  4. Open Resources and Plasmid Tools For Studying C. elegans

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    Blog Post
    ...elegans. To study the role of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a...polyadenylation for all protein coding mRNAs, and annotations extracted from other databases (Mangone et al...Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans...methods provides unique opportunities to develop computational methods to better understand animal development...additional SapTrap plasmids used for introducing point mutations that was published in a microPublication (Schwartz... E. M. "SapTrap vectors for introducing point mutations with unc- 119 + selection." MicroPublication 1...
  5. Hot Plasmids - October 2020

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    Blog Post
    ...you'll find in this post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein...corner New from the viral service Labeling excitatory and inhibitory synaptic proteins with FingRs ...developed AAV vectors to selectively visualize excitatory and inhibitory synapses in the brain.  ...scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively. Once expressed... fluorescent proteins targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers... development, regeneration, and certain disease states. However, it’s difficult to observe the cell cycle...
  6. How to Prepare for an Industry Interview

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    Blog Post
    ...Scientist positions often required a one-hour presentation about a prior research project. In total, the...notes after each interview.   Tailor your 1-hour presentation to your audience Start by asking the hiring ...hiring manager or recruiter what presentation format they require. Frequently they will ask you to present ...giving them sufficient background info. Use the presentation as an opportunity to demonstrate technical and... potential technical difficulties. Have your presentation on a flash drive and consider bringing your ...company will allow it. Before you start your presentation, introduce yourself to your audience. Pay...
  7. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

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    Blog Post
    ...efficient. Informed by these mutations and previously described TadA mutations (Gaudelli et al., 2017), they...withstand antibiotic selection. As a result, 8 mutations in TadA could create all three edits and were ...placeholder for the number that identifies the mutation in the evolved TadA domain. Because ABE7.10 contains...therapy approaches can use base editors to correct mutations in stem cells and return the corrected cells back...tested ABE8.17-m and found that one base change mutation (ABE8.17-m+V106W) (Gaudelli et al., 2020) was ...
  8. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

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    Blog Post
    ...replicate while maintaining the undifferentiated “stem” state and, Their potency or capability to differentiate... mostly from abnormal and/or supernumerary preimplantation embryos that are produced during in vitro fertilization...during ESC isolation. Low Chances of post-transplantation rejection when used therapeutically High Low...similar functional utility, iPSCs bypass the transplantation compatibility issues and ethical concerns of... used therapeutically, the chances of post-transplantation rejection are low. Further, as iPSC generation...embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis. Hum Reprod, 2008. 23(1):...
  9. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

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    Blog Post
    ... track of your mutation. To maintain stable lines of flies carrying a specific mutation, Drosophila geneticists...contributed by mother and father). While some mutations can remain stable as homozygous alleles, others...you can stably maintain a heterozygote, and the mutation won’t drift away with each passing generation....journey of trying to get males to recombine two mutations onto the same chromosome. You should use females...whether any phenotypes have been observed from mutations in the gene, and whether any publications have...
  10. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

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    Blog Post
    ...localization and transport between healthy and diseased states can also provide interesting insights into disease...analysis software produces quantifiable results and statistics that can be used to compare between different...Dunn et al. 2011 provides an in-depth review of quantitative colocalization analysis methods and useful image... image analysis software tools. Pitfalls, limitations, and specialized colocalization methods Fluorescent... very useful tools, but they are not without limitations and well-executed experiments require some consideration...Fluorescent Proteins. Ruusuvuori, Pekka, et al. "Quantitative analysis of dynamic association in live biological...
  11. PhD Applications After COVID

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    Blog Post
    ...challenges, and the flood of interview requests and visitation days can be, at times, very overwhelming. As ...applied to PhD programs in Biology around the United States in 2021, I know that this process only became even... to a school that requires its students to do rotations in different labs before selecting an advisor,...prospective student of such-and-such university State how you found their lab (usually through the university...name of the award you won for your conference presentation. It can help soothe your nerves and make sure...on opposite sides of the country had scheduled visitation days during my midterm exams. Using the online...
  12. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

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    Blog Post
    ...in an open inactive state, a closed active state, and an artificially active state due to closure by FP...The FP is confined to a plane and generally only rotates in 360°, increasing the probability that two FPs... and Citrine) can be monomerized with an A206K mutation [2]. Addgene has many of these constructs and ...used to determine if a protein is monomeric is sedimentation equilibrium analytical ultracentrifugation [...
  13. Genome-wide Screening Using CRISPR

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    Blog Post
    ... (NHEJ), usually results in a loss-of-function mutation within the target gene. CRISPR can also be used...cells, with each cell or set of cells containing a mutation in a different gene. Libraries may be available...correspond to genes that confer drug resistance when mutated. Findings from this type of experiment can shed...requires the use of next-generation sequencing. Representation - Most libraries contain 3-6 gRNAs per target... gRNA within the population is key. Loss of representation due to enrichment or depletion of specific ...CRISPR screen. However, maintaining sufficient representation within your mutant population requires a massive...
  14. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

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    Blog Post
    ...These insights are hard to appreciate without computational support. Indeed, rational sgRNA design is not...painstakingly assessed to something a bit more representative of the communities’ opinion. The trouble is...pool of CRISPR software embodies solutions that facilitate most experimental applications in CRISPR engineering...function can be found in their online tool’s documentation. Since Hsu et al., 2013, other tools have incorporated...target sites, off-target sites, scores and the statistics that go into calculating them. For this, some... hardware. This OpenCL dependency is a severe limitation, but the tool is fast enough that, if you’re ...Python script called SSFinder, but I find its implementation to be too slow for practical use. The R based...
  15. Tips for arabidopsis transformation

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    Blog Post
    ...Agrobacterium tDNA insertion likely precipitated domestication of sweet potatoes (Kyndt et al., 2015)! It sounds...plants. If your cultures have been incubated at stationary phase for too long, the cells in your culture...affected by the selection eventually. Genotype putative T1 transformants: When your T1 is on soil and .... et al. (2015) ‘The genome of cultivated sweet potato contains Agrobacterium T-DNAs with expressed genes... the National Academy of Sciences of the United States of America. National Academy of Sciences, 112(18...
  16. Getting to Know Expansion Microscopy

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    Blog Post
    ...inspiration behind ExM, how it works, and some of the limitations to keep in mind whether you are planning your...your specific targets. What are some of the limitations?  A “super-resolution” technique that uses common...great, right? But there is always a catch - some limitation of the technique that users should be aware of...First, expansion requires fixed samples. This limitation may be the both most obvious and least flexible...ensure the expanded images are still a good representation of reality. Third, expansion results in dimmer...density (Mikhaylova, et al. 2015).  Despite these limitations, ExM is a valuable tool, especially when paired...
  17. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ... Albert Einstein might have put it best when he stated, “the value of an education . . . is not the learning...Provost, then a new faculty member at Minnesota State (and now Professor of Chemistry and Biochemistry...Community (MCC) that seeks to reduce barriers to implementation. Funded by the NSF (with Co-PIs Jessica Bell...Professor and Chair of the Biology Department at Keene State College in New Hampshire. He mentors undergraduates... Kowalski, J., Hoops, G., Johnson, J. 2016. Implementation of a collaborative series of classroom-based...Scientific Elite: Nobel Laureates in the United States. Free Press. New York. Resources MDH CUREs Community...
  18. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

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    Blog Post
    ...development, sex determination, and anterior-posterior segmentation (Jennings and Ish-Horowicz, 2008). Fruit flies...flies that contain a specific loss-of-function mutation in groucho have thick tufts of sensory bristles...with non-functional hedgehog exhibit abnormal segmentation during embryogenesis. Typically, regularly spaced...embryo depending on its concentration. Genetic mutations in SHH cause severe health conditions such as ...reconstructed a transposase system based on the naturally mutated and non-functional transposase gene found in salmonid...Nüsslein-Volhard, Christiane, and Eric Wieschaus. "Mutations affecting segment number and polarity in Drosophila...
  19. Hot Plasmids Spring 2024

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    Blog Post
    ...screens Improved prime editor PE7 Antibodies to facilitate SARS-CoV-2 research New viral vector preps of...-024-07259-6. PMID: 38570691. Antibodies to facilitate SARS-CoV-2 research   By Ashley Waldron Ready-to-use...HaloTag, Figure 4), they determined that an A122D mutation in the rhodopsin domain conferred several new ...potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also...-microglia cells.      Figure 5: A) Representative confocal images showing GFP expression from...System By Vaibhav Kawde To circumvent the payload limitations of traditional AAVs, the Becirovic Lab has developed...
  20. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    Blog Post
    ...engineered by introducing Cas9-mediated frameshift mutations in the gene of interest. The genomic target for...for Cas9 and the introduction of this mutation can be any site early in the coding region of the gene ... experiments may require the introduction of a mutation at a specific site e.g., deleting the catalytic...modification at a defined amino acid to mimic a patient mutation or maybe to disrupt a catalytic residue. Good ...to either cytosine or adenosine deaminases to facilitate a single base pair edit. (Cytosine deaminases...of activators include the SunTag system which facilitates recruitment of multiple activation domains to...
  21. Changing Labor Laws Bring Increased Postdoc Wages

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    Blog Post
    ...under this ruling, regardless of visa or fellowship status, in addition to certain staff scientists and those... salary was set at $47,476 and the date for implementation was set for Dec 1 2016. In order to be exempted...paid. Institutional salary policies are one indicatator of how salaries may changes due to the FLSA. ... The University of Notre Dame sets no minimum, stating that while they recommend NRSA levels, postdocs...not to be tracked. University of Buffalo and Ohio State both have minima of $30,000, and raising salaries...addition, both Brown and Brandeis universities have stated that they consider postdocs paid on stipends to...
  22. Tips for CRISPR Gene Editing in Mice

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    ...how many pups will contain a mutation and many of the mice with the mutation will be homozygous for it in.../Cas9 for gene editing has expanded since its adaptation for use in mammalian cells in 2012-2013. Researchers...region. Or you may even wish to generate a point mutation and need to focus your attention to that particular...will require you to house mice that may have no mutation, costing money and space. As a rule, screen all... et al. "One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome...
  23. Important Considerations in Optogenetics Behavioral Experiments

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    Blog Post
    ...Garret D., et al. "Excitatory transmission from the amygdala to nucleus accumbens facilitates reward seeking...to your field. Ultimately, your question should dictate your behavioral output. However, regardless of ...behavioral test in which your circuit of interest is putatively involved in regulating the behavioral output....2) there is some problem with the technical implementation of your optogenetics setup (viral expression...and anxiety along the dorsoventral axis of the dentate gyrus." Neuron 77.5 (2013): 955-968. PubMed PMID...
  24. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ... with sub-optimal codon usage (Table 1) A few mutations are common to all or most expression strains to...levels including:  ompT: Strains harboring this mutation are deficient in outer membrane protease VII, ...methylate DNA. dcm: Similarly, strains with this mutation are unable to methylate cytosine within a particular...Tetracycline RecA-deficient; best for plasmids with repetative sequences.  F- ompT lon hsdSB (rB- mB-) gal ... of unstable proteins Tuner (DE3)   Contains mutated lac permease whch allows for linear control of ...Contains highly active thioredoxin reductase and glutathione reductase to faciliate proper folding; may increase...
  25. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...and CRISPR/Cas9 have also been used to create mutations in genes essential to HIV-1 replication. However...CRISPR therapies through PAM or seed sequence mutations. Multiple gRNAs could be used to combat this problem...LTRs or essential genes. Many of these escape mutations were located near the Cas9 cut site, leading Wang...escape occurs in an animal model. If these escape mutations are frequent, the dCas9-mediated “shock and kill... Zhang T, Li F, Yang W, Kaminski R, Fagan PR, Putatunda R, Young WB, Khalili K, Hu W. CRISPR/gRNA-directed... S, Wainberg MA, Liang C. CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 Replication and Accelerate ...
  26. Performing In Vivo CRISPR Screens Using the FITS Approach

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    Blog Post
    ...CRISPR screens in immune cells have identified many putative regulators of immunity. However, in vitro screens...approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation of modified cells; and (5) is compatible...molecular identifiers, which can improve the statistical power of in vivo screens. Figure 1: Using...used the pRDA_526 vector to knock out 30 genes putatively involved in CD8+ T cell responses to cancer. ...pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses in tumors...
  27. Keeping Up With CRISPR/Cas9

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    Blog Post
    ...that CRISPR's could be used to correct disease mutations, both in mice and in human cells. But that important...genes in one step without producing off-target mutations," Sha was quoted as saying when the news first...blogger Caroline LaManna put it well: "Wow, custom mutations in monkeys already. I know we've discussed it ...news was circulating. Scientists at North Carolina State, in the labs of Chase L. Beisel and Rodolphe Barrangou...structure may help us engineer around the current limitations of the Cas9 complex," said study author F. Ann...
  28. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ...Bicistronic vectors combine excitation and inhibition by using both excitatory and inhibitory channels activated...light within a specific range of wavelengths. Excitatory channels allow positively charged ions such as... the opsin.      Figure 2: Diagram of excitatory (ChR2) activation by light showing ions flowing...signaling cascade that may go awry in a disease state.      Figure 4: Example of an intracellular..., H. (2019). Light-activated chimeric GPCRs: Limitations and opportunities. Current Opinion in Structural...
  29. A Control for All Seasons

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    Blog Post
    ...positive and negative controls to ensure proper interpretation of results. A positive control is generally...the precision and accuracy and therefore the statistical power of an experiment. Technical replicates ...autofluorescence, also interferes with data interpretation and can be addressed with a label control. ...autofluorescence and should be considered in the interpretation of results. Include the label control for all...membrane. A loading control is critical for data interpretation, especially when changes in protein expression...Waters, Jennifer C. “Accuracy and Precision in Quantitative Fluorescence Microscopy.” Journal of Cell Biology...
  30. PiggyBac-ing Through the Genome Editing Field

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    ...and a transposase. The piggyBac™ transposase facilitates the integration of the transposon specifically...completely seamless manner, leaving no sequences or mutations behind. Furthermore, piggyBac™ offers a large ...achieved using the piggyBac™ transposase, which facilitates direct integration of the donor-transposon (carrying...standard homology directed repair template to facilitate the selection of cells containing your desired...excision-only piggyBac™ transposase (PBx), which contains mutations making it competent for excision, but defective...primarily two other ways to create specific point mutations by gene editing. First, instead of using piggyBac...
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