...variants differing in kinetics, affinity, and ∆F/F0. Introduce the two variants in three different membrane-targeting...fluorescent signals upon sensing glutamate in mouse and ferret models. Furthermore, they created three affinity...medium affinity for glutamate), that came in three different colors – blue (SF-Azurite-iGluSnFR), green (SF-iGluSnFR...glutamate signaling events, and accurately differentiating between synaptic vs. extrasynaptic glutamate...Figure 1   Increased versatility through two different variants The yellow (v82) and lime green (v857... Figure 2: Glutamate titration curves for the different iGluSnFR variants. Image adapted from Aggarwal...Figure 3: pH titration curves and ∆F/F0 for the different iGluSnFR variants. Image adapted from Aggarwal...

...can't distinguish the different proteins from each other. To distinguish different target proteins in your...target. By using a mix of different antibody-fluorophore combinations for different markers of interest, you...occur in combination with that reporter, you can infer its presence through detecting the reporter's fluorescence...a process called Fluorescence Resonance Energy Transfer (FRET).   Figure 1: Emission spectra and...commonly used fluorophores. A fluorophore's brightness refers to the signal strength of the emitted photons detected...a global maximum at around 560 nm, leading to different levels of brightness, depending on the laser light's...these aren't strict rules and in practice the differences in brightness levels are often not that great...

... in a different city, you can still develop a successful FRET-based assay since energy transfer only occurs...what's it good for? Fluorescence resonance energy transfer (FRET) is a biophysical interaction between two...first fluorophore (the donor) is excited, it can transfer energy directly via resonance to the second fluorophore...targeting the donor and acceptor fluorophores to the different biomolecules you’d like to test for interaction...that the targeting specificity requirements are different for the donor and acceptor fluorophores. The donor...Å away from the donor doesn’t result in energy transfer and thus, your specific FRET signal is unaffected...unaffected  Due to these fundamental differences in targeting specificity for donor and acceptor fluorophores...

...overexpress or knockdown a gene, you might need different plasmids. Harvard’s DRSC/TRiP Functional Genomics...varying expression levels of your transgene in different fly lines. Also, if there is a chance you need...transgenes in one fly, having the transgenes in different loci allows for genetic recombination so that ...for direct comparisons between lines expressing different variants of YFG because the transgenes are found...remember that you’ll need to integrate them all at different loci. Fortunately, there are many docking sites...docking site is located). This amounts to a small difference of two or three weeks. 3. How would you like ...yourself. Depending on your strategy and the services offered by the company, you might have to map into which...

...digestion be sure that the buffers and temperature are compatible between the different enzymes. Always check...ligation mix in different bacterial strains (stbl2, NEB stable, etc.), and/or use a different temperature ...want to assemble together in a combination of different ways according to your cloning strategy: Restriction...from other DNA fragments and you will remove the buffers used in the previous reactions. This will increase... There are multiple ways you can assemble the different parts of a plasmid based on the cloning strategy...attention in the areas of ligation between the different fragments If you don’t get any positive clone...might want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...

...current job situation. What can a coach do that is different from an adviser, boss or mentor? First, professional... transition out of the academic sphere into a different meaningful career in science. Supervisors in the...speaking at research institutions and at scientific conferences, I see that few scientists are aware of the benefits...few hours of coaching time can make an enormous difference so don’t assume that this will cost a fortune...successful outcome. How do I find a coach? Start with referrals. Are there scientists you know who have had a ...offices or professional organizations can provide referrals. There are bad coaches out there, so finding a...specializes in scientist coaching. Always ask for references and follow up with 3-4 to hear about those clients...

...containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral elements...it is recommended that you try a few different ratios of transfer, envelope, and packaging plasmids in...associated with producing and using lentivirus and offer some tips and tricks for overcoming these hurdles...transactivator, Tat. For more information on the different generations of lentiviral packaging systems please...Guide. While the third generation packaging systems offer an extra level of safety, they also require a fourth...encoding the structural elements, envelope, and transfer gene are transfected into a packaging cell line...titer. Importantly, optimized conditions for one transfer plasmid are not always the best for others. For...

... Evaluating Gene function by RNA interference (RNAi) RNA interference (RNAi) is a powerful tool for performing... These smaller dsRNA fragments are referred to as small interfering RNAs (siRNA) and they bind to a class...genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to knockdown expression of specific gene...morgani, and Phagocata gracillis. There are 11 different genes cloned into plasmids to investigate for ...WISH technique. The expression patterns of 11 different planarian genes (all of which are available through...one is working with, the protocol will slightly differ. For example, Girardia sp. and Dugesia dorotocephalacan...and if you make new plasmids for investigating different planarian genes, we encourage you to deposit them...

...fear of referencing a wrong section. The antibody screening and development of a focused reference bring...School of Medicine at Mount Sinai.  References and resources References Baker M. Reproducibility crisis: ...), variability (separate batches can perform differently – happens when the antibody is produced from ...a new set of animals), and wrong application (different experiments/conditions can change binding ability...off-target molecules), reliable (wouldn’t perform differently over time), compatible with the desired applications...and accuracy while screening. We also compare different clones of the same target to see if the results...with general non-specific binding. If the sample differs from the negative control, such as showing a very...

...for contributing their expertise! References and resources References Aschauer, D. F., Kreuz, S., & Rumpel...Stable Pancreatic Gene Transfer by Adeno-Associated Virus Vectors via Different Routes. Diabetes, 55(4...multiple coreceptors, which may be present on different cell types and sometimes vary between target species... numbers separated by a slash: the first number refers to the genome, and the second number to the capsid... al., 2021 In addition to tissue tropism, different AAV serotypes can be useful to evade neutralizing... (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1, AAV6, AAV7, and AAV8 in the ...., Yin, L., Merigan, W. H., Flannery, J. G., & Schaffer, D. V. (2013). In vivo-directed evolution of a...

...frequently implicated in cancer cell proliferation, survival, differentiation, and apoptosis. For each of these...controlling properties like growth, survival, and differentiation.” https://www.addgene.org/kits/sabatini-wood-cancer-pathway-orfs...cells one at a time, you could identify those that confer resistance to drugs,” in a rapid, straightforward.... “You could use it to screen for pathways that confer any phenotype of interest, not just drug resistance...morphological features, migration, or survival in different contexts.” It’s also possible you could make use...plasmids in living animals, to explore pathways that confer metastatic potential or the ability to survive ... sometimes drug screens and sometimes totally different experiments we never thought of - and that’s thrilling...

...the non-profit sector are transferrable to industry such as technology transfer, communications, medical...skills overtime for success. Many people will have different roles, titles, and responsibilities during their...candidate is ultimately looking for a role in a different functional area, then the research role could ...higher degree. These numerically limited visas are referred to as “cap” subject H-1Bs. The H-1B allows U.S...and therefore the academic H-1B visa cannot be transferred to an industry position. If you cannot get an...a foreign national to have an employer or a job offer in order to obtain permanent residence. The recently...your field. No employer is necessary and no job offer is needed for this category. To qualify, you need...

...cellular proliferation. We have appreciated for some time the causes of this uncontrolled proliferation, in...normally encourage cell survival, growth and proliferation, while tumor suppressors perform the opposite...or, in the case of cancer, uncontrolled cell proliferation. Hence it is imperative to the health of an ...division. The combined effect is unregulated proliferation. Another safety net is our own immune system...cellular pathways involved in the development of different cancers and identifying the roles of key proteins...is the Ras pathway. The term “Ras” collectively refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded...approaching the Ras problem by exploiting the different mutant versions of Ras that are most prevalent...

...presenting your research at conferences such as SACNAS and ABRCMS. These conferences have a lot of programming... recruiters will attend these conferences to answer questions and offer application fee waivers. One of...her on Twitter @EleanorWang.     References and resources References Canning EA, LaCosse J, Kroeper KM...neurodegenerative disease because they had close relatives suffering from similar diseases. Others were inspired by...proactive Rather than applying only to posted job offerings, the best research experiences typically come ... is a supportive environment—it makes all the difference in a lab experience. “Mentorship” was a bit of...interested in doing research and attending research conferences. The Barry Goldwater scholarship is also a national...

...primary antibodies raised in different species, or at least from different IgG subclasses, so that secondary...incubation of 1° and 2° Must use different species and/or different IgG subclasses of 1° for multi-color...limited cross-reactivity between different IgG subclasses and different species, nanobodies are useful ...Multiple polyclonal secondary IgGs can bind to different epitopes of a primary antibody. B) Each arm of...they don’t require using primary antibodies from different IgG subclasses. Instead, you can pre-incubate ...Once you know the answer to these two questions, refer to this table to find the right nanobody for your...come in 1x and/or a 3x cysteine varieties. This refers to the number of cysteine sites the nanobody has...

...capture the fact that different parts of the world encountered COVID-19 at different times. Most notably...plasmid requests Figure 5: Inferred effects. Values are the inferred percentage of normal activity ...scientific fields could be different.  Conclusion Figure 7: Mean inferred effects by region. In...also eases the interpretation of the parameters we infer, since scientific productivity is better interpreted...regions. Each of these regions was affected at a different time by COVID-19, so we defined the COVID-19 event... day) on dates impacted by each effect. The inferred parameters suggest that COVID-19 induced a 2x–...Europe most impacted and China least impacted. The inferred weekend effect is, in retrospect, generally more...

...serve similar purposes, both techniques offer researchers different advantages and limitations.  The four...factors in which chemogenetics and optogenetics differ are: Timing Targeted manipulation Controlling ...study. Figure 1: A schematic showing the differences in chemogenetics and optogenetics in a mouse ... light (optogenetics).   Timing Optogenetics offers a high degree of temporal resolution as receptors...neurons to repolarize faster.  Chemogenetics does not offer the same level of control over timing. Because chemogenetics...amounts of stimulation to receptors. Optogenetics offers better control over these factors as the light ...manipulated or turned off. In chemogenetics, while different concentrations of ligands can be injected, this...

...going to cite hundreds of references) and it’s better to keep your references organized from the beginning...sense of the similarities and differences among proteins in the class. Offer your perspective It doesn’t... tool kit. I wrote a review article on histone modifers in B and T cells with my adviser when I was a ...criteria to shape the document. Get and use a reference management program (e.g., EndNote, Papers, Mendeley...Mendeley, etc.) You’re going to be managing a lot of references. I cite as I write, meaning I use the software...in real time as I wrote, so each statement was referenced even in the roughest version of the draft and...that people can go back and read the original reference for the data. For example, we summarized how frequently...

...methods can interfere with binding, and some common antibody buffer components can interfere with conjugates...scientists have since adapted it for a host of different proteins and small molecules from a variety of...needs. Herein, we’ll give you a rundown of the different types of ELISA, advantages and disadvantages of... non-overlapping epitopes so that they do not interfere with each other. The detection antibody may either...this method, the target antigen competes with a reference antigen for binding to a primary antibody.  While...typically users first coat the multiwell plate with a reference antigen and then they preincubate the antigen-...allow the unbound primary antibody to bind to the reference antigen. The more antigen in the sample, the more...

...with that…and happy flowing!    References and resources  References Alyamani, A., Kalamegam, G., Ahmed...usually tell where this point is by looking at the different populations. In the case of Figure 2, one population...   Figure 3: It's often easier to identify different populations by imagining (or drawing) a quadrant...population and everything outside the gate is a different population.  Gating and plot orders  As the gates... right.  First, SSC and FSC are measured to differentiate cells from debris. Cells are gated, and in the...dendritic cells is further separated into three different populations using conjugated antibodies against...help me as a reader if I'm struggling to see the different populations on a plot.      Figure 5: ...