...assume that their advisor alone applies for these competitive grant applications; however, in some cases, your...the grant application process. When my PhD advisor first asked me to write a grant application due the ...mind that universities require application submission in-house approximately 2 weeks before the RFA due date...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...writing, volunteered to take the lead on various applications throughout my graduate career, and helped receive...opportunities to convince others why a project is the best approach to a given problem, Empowers the writer, And...

...intended application - Remember, just because your antibody performs as expected in one application does ...Strategies This approach compares target expression of antibody-independent approaches to your antibody-dependent...Genetic Strategies This approach uses a gene knock-down or knock-out approach to show that an antibody...protein. This approach is ideal for validating antibodies for immunocapture applications and is amenable...pause and make sure that your antibody has been appropriately validated so that you can have confidence in...in your results. But what is appropriately validated? This post will go over some of the general considerations...strategies for antibody validation in different applications. Here, I want to highlight the general suggestions...

...allow you to approach that problem in a novel way. Finally, engineering new applications to solve problems...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...someone else in the department) to read over your application and give you feedback. They’ve most likely been...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be ...

...of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy has a region of...directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the gene delivered...  3. Hybrid  Overview: The hybrid approach combines the overlapping and trans-splicing methods. The AAV...has a slight advantage over the overlapping and trans-splicing approaches since there are two ways for a... genomes is the basis of several split vector approaches developed in the early 2000’s which allow researchers...is not yet known. Drawbacks:  Generally an overlapping split vector has equal or higher transgene expression... on the rate of transgene expression with an overlapping AAV since both of these variables influence whether...

...is this application - the creation of sequence-specific knockins - for which the AAV approach is most ...for a given experimental approach. First, unlike CRISPR, the AAV-based approach is only able to target ...simultaneously in the same cell, are more appropriate for a CRISPR-based approach. Second, the AAV preps generally...limitations, the AAV approach is best suited for tissue culture based approaches that require modification...researchers could apply this technique to efficiently deliver large repair templates for applications that require...working to develop high efficiency gene targeting approaches improved on this finding with the addition of...challenging to develop high efficiency CRISPR-based approaches for the generation of knockins via homologous...

.... This is the direct approach to immunoimaging assays. But if the direct approach isn’t giving you enough...   Figure 1: The direct approach (left) versus the indirect approach (right). Primary antibodies ...varies across applications. Western blots, for instance, almost always use the indirect approach, while flow...immunocytochemistry may all use either the direct approach or indirect approach, depending on the requirements of the...interest, it may be time to look at the indirect approach - which means it’s time to talk about secondary...binds to the primary antibody. In the indirect approach, the signaling molecule is conjugated to the secondary...antibodies can bind to a single primary antibody, this approach can greatly increase the sensitivity of an assay...

..." are summarized below. Check Out Erik Snapp's Full "Applying for a Faculty Position" eBook Here 1. Have...  Erik Snapp just released version 3.4 of his eBook, How to Get a Faculty Position. This edition has ...along with new thoughts on research statements. Snapp, who is always looking for ways to improve his eBook...suggestions, please do not hesitate to email me at snappe@janelia.hhmi.org." You can find his original blog...years ago, I decided to write a "how to" manual on applying for faculty positions in biomedical science. My...talents and ideas. Some key points from my ebook, "Applying for a Faculty Position: the View from Both Sides...experts in your field. Not true. Even if you are applying to a department in your specialty area, it's a...

...safe for clinical applications. Excisable Cre-lox lentiviral vectors: This approach is the same as using...of excision, this approach is still not considered safe for therapeutic applications. piggyBac (PB) transposon...This is considered a safer approach for developing therapeutic applications, but it can take a long time...time, scientists have used almost all available approaches for generating iPSCs. The generation of iPSCs...Retroviruses are not considered safe for clinical applications.     Lentiviral: Lentiviral vectors infect dividing...dilution via cell division. A drawback to this approach is it has lower levels of efficiency at generating...express reprogramming factors for therapeutic applications. Sendai viral vectors: Sendai virus is a single...

...relationships. However, most job seekers will and should apply for jobs posted online as one part of their job ...Utilizing existing relationships as part of the job application process can dramatically increase the chances... of a resume can be crucial for a successful application. It takes time to prepare an excellent resume... Many job postings will result in hundreds of applicants the day they are posted.  If your resume is not... here for more on timely submission of a job application. 3) Read job descriptions carefully Be open minded...have to have all of the listed qualifications to apply for a position.  A job description is usually a ...wish list and the hiring manager knows that no applicant is likely to meet all the criteria. Note the criteria...

...skipping approach in mice, DMD researchers are very excited. This approach may also be applicable to a number...success in basic research, researchers are eager to apply it in a clinical setting. CRISPR is often used for...), an X-linked recessive disorder affecting approximately 1 in 5000 males. DMD is caused primarily by ...very large (79 exons), but much of the sequence appears to be nonessential. Within the mutational hotspot...so researchers have set their sights on another approach. Since shorter forms of dystrophin can still be...researchers have begun to explore genome editing approaches for exon skipping. Addgene depositor Charles ... removal rate of exon 51, which resulted in appropriately localized dystrophin. In a subsequent study,...

...genes provided by the user. This approach is useful for several applications as will be discussed below. The... screen approach is to use smaller libraries targeting subsets of genes. With such an approach, you lose...custom CRISPR screen and the appropriate phenotypic readout, similar mapping could be rapidly performed ...challenges with a custom screen approach is to identify an appropriate list of genes to target. Thus, ... involved in a studied cellular behavior. This approach is particularly suitable for in vivo screens, ...drugs. Known drug targets  Another interesting approach is to design screens against known druggable targets...preferentially those for which validated inhibitors or even approved drugs already exist. If you are lucky and find...

... tools for familiar applications. But how does one invent a tool for applications that don’t even exist...and develop potential applications.  Andrew is a frequent collaborator and is happy to give advice and support...phenomenon, improve the tool, and develop new applications for magnetoresponsive proteins. Motivated by...organisms) are opaque, which makes most biomedical applications of these tools difficult or impossible. But ...tool with magnets would enable new research and applications that don’t yet exist, but it wasn’t clear that...molecules like proteins with magnetic fields generally appears to be weak — if they exist at all. Andrew and Maria...fluorescence intensity decreased whenever the magnet approached the sample (by less than 1%) and recovered after...

...single model. The first approach can lead to new directions, while the second approach could terminate with... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia.... Given this large sum, one might expect that applicants would work for months to hone a chalk talk. Yet...preparing for both types. Your research vision is your approach to resolving a significant knowledge gap in your...has unclear utility. You cannot predict what will happen. Without the prediction, you have no hook. A battery... you need to be able to describe experimental approaches in terms of the question, the methods, the tools...

...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular... post was contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the... of a mystical "Force," this quote is equally applicable to many microscopy experiments with fluorescent...not fluoresce. A dark population is not readily apparent and can confound quantitative imaging experiments...for use in cytoplasmic environments. However, approximately 40% of human (and most eukaryotic) proteins ...have reported that FP cysteine residues form inappropriate disulfide bonds in the secretory pathway (Figure...localization. For example, the formation of inappropriate inter-chain disulfide bonds between FPs blocks...

...developed and applied. The current pandemic serves as a striking example of the applicability and importance...research. I applied, hoping that the experience would improve my scientific and application writing skills...science (as opposed to translational, biomedical, or applied sciences), people conduct research to answer questions...revelation. Science in any field can generally be applied to improve our lives and solve global problems....grants Varying degrees of engagement with what happens in the lab Official title is typically Professor...the people around you. Be proactive Rather than applying only to posted job offerings, the best research...ties into the previous one too. I would recommend applying for multiple positions—don’t put all your eggs...

...received 116,224 employment based green card applications and approved 91,023 during the first 3 quarters of ...options discussed in this post benefit from the applicant thinking about the process early, keeping good...is a numerical quota of 85,000 total H-1B visas approved each fiscal year: 65,000 visas plus 20,000 reserved...occupation" as requiring theoretical and practical application of a body of highly specialized knowledge in ... April 1 deadline for companies to submit an application on their behalf.  If an individual obtains a ... cap subject H-1B (last year less than 30% of applicants received sponsorship) then he or she can start...1B petitioners are exempt from the cap if the applicant works at institutions of higher education, a related...

...real-time PCR (4). Restriction mapping and modern sequencing Restriction mapping informs on DNA sequence through...useful applications and have informed many current techniques. For example, restriction mapping led to...historical techniques: southern blotting, restriction mapping, and sequencing gels, as well as their modern equivalents...comparison to a control sample. Southern blotting applications range from confirming the results of cloning...gene that causes sickle cell anemia. The mutation happens to destroy a sequence recognized by the restriction... was commonplace, researchers used restriction mapping to investigate DNA sequence content by comparing...of the DNA. Starting in the 1970s, restriction mapping was commonly used to map DNA and examine genomes...

...zebrafish embryo has been mapped using single cell RNA sequencing approaches (Harland, 2018). Several ...investigations. Genetic resources for zebrafish A common approach to unravel the gene networks that drive biology...DNA into the genome is useful for a variety of applications. For example, specific tissues such as blood... regions is also possible using an alternative approach that exploits the bacteriophage integrase phiC31...useful, I’ll outline two recent lineage tracing approaches that led to important insights in developmental...tracked using microscopy and advanced computational approaches (Keller et al., 2008). This microscopy-based ...pathways control this process. Additional zebrafish applications and conclusions Zebrafish are also popular outside...

...subsequent, repeated activation), this is an approximation of reward or the sense of pleasure that the ...EthoVision. Consult the literature to find all the applications of optogenetics relevant to your field. Ultimately...behavior, certain technical considerations still apply. Testing stimulation First, In lieu of electrophysiology...stimulate them for 30min to an hour and observe what happens. You never know how stimulation of a particular...optimization, turn on the laser and watch what happens. Who knows, you may observe something that could... discovery. Monitoring behavioral output The appropriate behavioral readout for your optogenetics experiment...brain region that may be important in memory, an appropriate behavioral test might be contextual fear conditioning...

... a bioinformatics approach examining genomes possessing CRISPR systems with apparent self-targeting. Two... plasmid or group of plasmids' functions and applications. We hope that these articles make it easier ...includes four promoters, six fluorophores with nonoverlapping emission spectra (N- and C-terminal tags for...Targeting and manipulating interneurons using a viral approach Genetically modified organisms like transgenic...successfully targeted via viral particles, an effective approach to target specific neuronal subtypes was not available..., Gordon Fishell’s lab has developed a new AAV approach using regulatory elements that restrict expression...tools in virtually any vertebrate. This novel AAV approach allows you to target and functionally manipulate...