We narrowed to 671 results for: FIE

...individual’s work in their field and the impact of the individual’s past work on the field. NIW requirements stress...in the field. This criterion is best proven through letters of support from experts in the field. You must... of the work of those in your field. Original contributions to the field that have had a major impact....specific academic field, (2) that you have three 3 years of research experience in the field, (3) that you...academic field Evidence of original scientific or scholarly research contributions in the field Evidence...expertise and impact on your field from the point of view of experts in your field. Please remember, this is...research role could be the point of entry into that field. In this post, we discuss the following paths to...

...protein tool that responds strongly to magnetic fields (Frank Hayward et al., 2024). They deposited their... you’ve ever had an MRI, you know that magnetic fields can pass right through our tissues unimpeded. Andrew...on whether and how organisms respond to magnetic fields (Karki et al., 2021; Kattnig et al., 2016), and...Wheeler et al., 2016; Duret et al., 2019). But the field has sometimes seen controversy, and the interactions...of organic molecules like proteins with magnetic fields generally appears to be weak — if they exist at.... Gray regions indicate times when the magnetic field is on. Image reproduced from Frank Hayward et al...magnetoresponsive system that could be expressed and purified. Although a ~1% change in signal might not make...

...fused to epigenetic modifiers enabled epigenetic modifications at a user-specified locus. Researchers...epigenetic modifiers available from Addgene. For most of these constructs, catalytically dead modifiers are ...this purpose. Epigenetics began as a correlative field in which covalent modifications to DNA or histones...fusions between Cas proteins and the epigenetic modifier could be targeted to specific DNA sequences. Non-editing...catalytically dead dCas9 fused to a variety of epigenetic modifiers to specific loci without inducing double strand...increases levels of H3K27ac histone modification at specified loci. Charles Gersbach’s lab has deposited mammalian...transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, is available from Rudolf Jaenisch...

...entire kit; a parallel analysis that successfully identified new, highly specific agonists for orphan GPCRs...Densmore’s lab at Boston University have greatly simplified the process of generating multiple E. coli gene...multiple screens in different human cell models and identified ~4-5-fold more fitness-related genes than have...across multiple species Recent discoveries in the fields of genome engineering and synthetic biology have...vectors that allows one to use the MAGE method in unmodified bacterial species (4). This set of plasmids (...oligonucleotide integration, allowing MAGE in otherwise unmodified bacterial strains (5). MutL is highly conserved...modifications has been to use inhibitors of epigenetic modifiers like DNA methyltransferases or histone deacetylases...

...exactly what aspect of the field you plan to review. If it’s a well-researched field, you may need to get specific...moving - remember you're becoming an expert in your field and are the perfect person to be writing the review... of exposing you to relevant researchers in the field. Finally, you can sound credible and cool when you...because, as I mentioned, I wasn’t an expert in the field. To find reviews, I just searched online and found...these articles to get a sense of the themes in the field and to learn what people cared about. I also used...to read. Once I had an idea of the themes in the field, I searched for recent research papers on these ...articles from the active/well-known researchers in the field. I made sure to find information from genome-wide...

...microscopy techniques   Fluorescent Widefield Microscopy Fluorescent Widefield Microscopy is the most common...to solve two drawbacks of widefield and confocal microscopy. First, widefield and confocal microscopy project... limiting photobleaching. Similar to widefield, the entire field of view is excited at one time and captured...objective evenly illuminating the entire (wide) field of view. Fluorescent light traveling back towards...samples are thin enough to sit within the depth of field of the objective (the distance in the axial dimension...these experiments are performed on a fluorescent widefield microscope. Here, excitation light is projected...projected from the objective so that the entire field of view is bathed with an even illumination. The emitted...

... further modified sdChr- a blue-shifted channelrhodpson from freshwater green alga identified by Ed Boyden...or Zinc Finger Nucleases, the Gersbach lab has modified the RNA-guided CRISPR-Cas9 system to create a ...lab have teamed up and advanced technology in the fields of single molecule imaging and inducible gene expression...by scFv antibodies fused to sfGFP. This system amplifies the intensity of fluorescence signal and enables...under the right conditions, and could initiate specified cellular processes or be observed at a later time...screening applications. The system can be readily modified to use different antibiotic resistances, restriction...gene of interest. The gene to be expressed is amplified using a gene-specific forward primer containing...

...lens which magnifies the image, then the ocular lens, where the image is further magnified before it finally...4x objective magnifies the image four times the actual size, the 10x objective magnifies it 10 times, ...also 10x, you would actually be seeing a 100x magnified image of that object! Equipment Light Microscope...aperture are in focus. Center the aperture in your field of view and then slowly open the diaphragm until...the stage (and ultimately your slide) until your field of view is centered on a region of interest. If ...

... the purified viral vector solution (which contains viral capsid proteins) as opposed to purified DNA....transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type AAV2 has... genome-containing viral capsids is typically quantified by electron microscopy of a viral vector solution...negative staining. Empty vector particles can be identified after negative staining and appear darker than...and comparability because each sample with be quantified with a different primer pair. Ct values vary...DNA. This may be mitigated by using purified viral DNA as the template instead of the intact AAV particle...AAV SFM), which is an AAV sample that has been quantified by 16 labs around the world and can be used by...

...necessary to remove iodixanol prior to use of your purified virus. Nonetheless, we recommend performing a ...a buffer exchange before using the purified AAV in vivo . The iodixanol gradient in this protocol is composed...genome containing) particles. On average, iodixanol-purified AAV preparations contain ~20% empty particles....60% iodixanol step Carefully add up to 5 mL of clarified supernatant on top of the gradient. Use 1X PBS...fractions obtained from the 40% phase contain the purified AAV. Option #3 Puncture the QuickSeal tube slightly...vertical black line indicates the location of the purified AAV. Left image adapted from Zolotukhin, S., et...the position of the needle for harvesting of the purified AAV using option #1. Right image adapted by permission...

...different fields is quite vast. If you’re a novice neuroscientist looking to get into the field, this is...neuroscience newbies might want to get into the field. Find Optogenetics Plasmids for Use in Neuroscience...Neuroscience Research Neuroscience covers many subfields When I first began looking into the details of SfN17...plenty of interaction between neuroscience and other fields. For instance I met one researcher studying how...right kind of biological expertise to get into the field, there’s likely some way you can use the tools and...from Addgene are already having an impact on the field Not to toot our own horn too much, but many people...find tools that will make your transition into the field much easier! If there’s a plasmid or AAV tool that...

...Microsoft Excel or similar software Reagents Purified antigen Purified recombinant antibody Isotype control antibody...describes how to run an indirect ELISA against a purified antigen to demonstrate the expected antibody dose...Enzyme-Linked ImmunoSorbent Assay (ELISA) against a purified antigen to validate an antibody. This protocol...the Antigen Standard and coat the plate Dilute purified antigen to 20 ng/µL in a microfuge tube and vortex...concentration. Prepare serial dilutions of the purified antigen as follows: 2 ng/µL : Add 100 µL of 20...coated with serial dilutions of human recombinant purified Desmin (blue) or a negative control protein, Apolipoprotein...

...healthcare industry publications like FiercePharma, FierceBiotech, and BioCentury can be an engaging way...Sachdeva. Healthcare consulting is a fast-paced field, requiring people who are willing to quickly learn...consultant and give you an idea of what a career in this field involves, and why you might enjoy doing it. I’ll...consultant and what you could do to break into the field. What is healthcare consulting? Healthcare consultants...communication skills is a must to be successful in this field. Personally, I have greatly benefited from and appreciated...healthcare economics, or in some combination of these fields. I got my PhD a couple of years ago in Bioengineering...you for a successful career in several different fields. While some people continue to enjoy the challenges...

...this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also contain reverse transcriptase...transcription and Cas13 is then used to detect the amplified region of interest. Cas13 uses a complementary...resulting in fluorescence. Cas12a binds all the amplified target regions and results in the continuous cleavage...subject to reverse transcription. This is then amplified by PCR using Taq Polymerase and then transcribed...Highlights: Reverse-transcribed RNA samples are amplified by PCR instead of RPA. Unlike RPA, this technique...typical PCR machines, the authors use affordable, field-ready, Bluetooth-enabled PCR machines that can even...made accessible by the DIY-Bio movement. Rapid, field-deployable nucleobase detection and identification...

...efficiently amplified than larger constructs, but plasmids as large as ~6 kb can be amplified fairly easily... the desired mutation) in a PCR protocol that amplifies the entire plasmid template. The parent template...obtained with PCR screening using the primers specified by the arrowheads in (A). Experimental guidelines...Methylation deficient bacterial strains can be identified by the dam13(-) mutation – you’ll want to steer...mutagenesis. Nuclease To remove the template DNA (unmodified plasmid) a restriction digest with DpnI is used...the bands present in the digest products of the unmodified plasmid will be split into two smaller bands ... two smaller bands visible in a digest of the unmodified plasmid into a larger band (not shown). As a ...

...different than what was originally intended. Sharing unverified plasmids can lead to far reaching consequences...sequences and/or features. Most of these regions are identified during the QC process, following sequencing and...confidence. If this approach does not resolve the identified differences, the plasmids are sent out to Plasmidsaurus...Illumina MiSeq.  Resolving QC issues If the QC team identifies a QC issue, they will contact the depositing ...depositing lab to review the discrepancies. Often, the identified issues will not affect function and the plasmid...page to make requesting scientists aware of the identified discrepancy in the plasmid, and that it is not...function, we request the lab provide a new, sequence-verified replacement for the plasmid, and the full QC process...

...regions that are bound to modified proteins called histones. We can detect modified histones with “GPS for...fractal). Enhancer biology is a complex and dynamic field. We’re going to focus on a tried and true method...DNA’s 3D geometry and the chemical properties of modified histones, a genomic region that has mono-methylated...genome math, we’ll need a bit of shorthand from the field of epigenomics: H3 – one of NANOG’s associated ...genetically programmed to CRISPR out the enhancers we identified in step 2. The result is a heterogenous population...enhancers, promoters, insulators, and more. I then identified valid gRNA sequences without off-target effects...terminator), and no off-target effects for the identified 23-mer gRNA. Off-target determination was established...

...library, followed by DNA isolation. The amplified library is verified through NGS sequencing to confirm library...Producing Your Lentiviral Library Once you've amplified–and verified!–your pooled library DNA, the next step...you’ll need to develop your own protocol that amplifies without disrupting plasmid representation. A good...backbone.  Verification Once your library has been amplified, you'll need to verify your library by next-generation... sequencing (NGS). The representation of your amplified library should be very similar to that of the ...libraries, NGS is the best way to validate your amplified library and verify that you have not skewed the...

...signal from this reporter can be measured and quantified to determine the presence and quantity of DNA...Enzymatic Reporter UnLOCKING Cas13 (C2c2) was first identified in 2016 by the Zhang lab as an RNA guided RNase...assay, targeted DNA or RNA from a sample is first amplified using RPA (recombinase polymerase amplification...RPA is coupled with T7 transcription to convert amplified DNA to RNA for subsequent detection by Cas13. ...used easily and effectively in the lab and in the field (Myhrvold et al., 2018). Now, the Zhang lab has ... SARS-CoV-2 RNA. The test is started using RNA purified from patient samples and can be read using a dipstick...reaction. To enhance sensitivity, the DNA is first amplified through isothermal amplification by RPA. When ...

...the CRISPR toolbox. We'll cover how Cas13a was identified, the structure and function of Cas13a with a ...cleaving CRISPR nuclease Cas13a was originally identified in 2015 (Shmakov et al., 2015). They were using...the bacterial genome. From this analysis, they identified 53 potential candidate genes that fell into 3...recognized and cleaved its target RNA sequence as specified by the crRNA sequence, it adopts an enzymatically...nucleotides containing a sequence of interest (red) is amplified using Recombinase Polymerase Amplification (RPA...Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex...enzymes continue to make a splash in the CRISPR field. We feel that the culture of sharing within the ...