...resulting in a decrease in luciferase activity. This experimental protocol can readily screen miRNAs for ...Peak emission (nm) Size (kDa) Substrate Cofactors Secreted Luminous shrimp(Oplophorus gracilirostris) 455...cell membrane and deliver the substrate; however, secreted luciferase or the use of alternative substrate...pathways can be investigated in a high-throughput screen using luciferase activity as a measure of signaling...

...that approach seems to create competition for your own spot. True! But you create a network of friends,... and simplify access to academic resources, we created STEAMid, for a nationwide database of  STEAM internships...with your question(s). This precautionary step increases the likelihood of acceptance to the internship... following graduation. While these suggestions increase your competitive edge, they alone, are no guarantee...

...the potential for increased speed and efficiency through which knowledge is created and shared. Can you...platform. Expanding further beyond the US and increasing Addgene’s international presence would potentially...information, making it available to everyone, that creates more breakthrough and innovation. With any kind...science regulatory law. My role at Suffolk is to help create new programmatic content as well as help our students...

...sequence desired. This technology allows you to create a pair of designer proteins that will introduce... transfection or electroporation. All you do to create a protein that binds those 15 base pairs is take...genome editing tools allow you to do is greatly increase the efficiency of the genome editing process. ...for investigators without a lot of expertise to create zinc finger nucleases that will bind desired sequences...in terms of the target site. Also, any time you create double-strand breaks, there is a possibility of...

...antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to...is a ~150 amino acid, single domain antibody. To create their RANbodies, Yamagata et al used previously... RANbodies are expressed in 293T cells and are secreted from cells due to their IgGκ signal peptide sequence...them. HRP RANbody staining results in a ≥10-fold increase in signal over staining with a primary/ labeled...

...Institute has created hundreds of thousands of plasmids, using both mass production genome wide screens and the...vector. The time it takes to make a plasmid could increase dramatically if you are piecing together several...at Addgene, the consumables cost per stab has increased 50% since 2019, while overall inflation of USD...equal to the $155 spent on reagents. Five ways to decrease plasmid production time and cost If you’re short...

...scaffold to enhance complex assembly, thereby decreasing the amount of background fluorescence from unbound...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...was fused to a different fluorescent protein to create three distinct colors. The specificity of these...distinct hairpin sequences produces white light. Created with BioRender.com. With this rainbow of colors...combination, for example, with a far-red FP, would increase the available colors to 15! They also envision...

...visibility is increasing internationally, thanks to evolving outreach tactics, increasing business development...they see the value in depositing and they are increasingly seeking us out on their own to share reagents... We were and remain ahead of most companies in creating a welcoming, inclusive workplace for everyone....about their dedication, excellent intentions, or incredible drive to succeed.  Thanks to all the Addgenies...

...address this need, the Khavari lab at Stanford created the RNA-protein interaction detection (RaPID) method...RaPID. Ramanathan et al. also tweaked BirA* to increase its rate of biotinylation. The improved enzyme...in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels by 90%, suggesting that ...Hyperferritinemia-Cataract Syndrome (HHCS), a disease characterized by increased iron serum levels and early-onset cataracts, is...sequences and that loss of binding correlated with increased serum iron levels seen in HHCS patients. These...

...also underwent editing through the NHEJ pathway. Created with BioRender.com. In later characterization...current tools, thus quieting some fears about the creation of “designer babies.” The differences in editing...disorders, pre-implantation genetic diagnosis (PGD) screens embryos to find those that do not carry a given.... claim that CRISPR would still be helpful in increasing viable embryos, especially in the case of older...still be worthwhile for a given disease if it increased the percentage of disease allele-free embryos ...

...Foundation, and Stanford University—that harnesses the creative power of sex and gender analysis for innovation... in research Governments, universities, and, increasingly, corporations in the US and Western Europe have...decades: “Fix the Numbers of Women” focuses on increasing the numbers of women participating in science...gender analysis and 2) provides case studies with concrete examples of how gender analysis leads to discovery... Environment—a project aimed at harnessing the creative power of sex and gender analysis for innovation...

...these demands? Are there other solutions, such as increasing numbers in Master’s level programs? What would...Maryland College Park, is a strong supporter of increasing the number of postdoctoral scientists at University...School. He has continually promoted the cause of increasing data collection about the scientific workforce... Boston 2015 where we hope to begin developing concrete solutions to the problems looming over the academic...School. In the lab, he uses synthetic biology to create tools to better understand human disease.  Outside...

...particularly given that this has precipitated increases to the NIH recommended pay scale – used by many...while this is a move in the right direction, the increase notably still does not meet the suggested postdoc...this area, keeping the pressure on for further increases will be challenging, but will be aided by the ... new strategies designed to cope with current increases. These two rulings along with progress over the...School. In the lab, he uses synthetic biology to create tools to better understand human disease.  Outside...

...grandsons that decreased their likelihood of death from cardiovascular disease but increased their risk of... DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG islands in the... units of the FRET pair together and causes an increase in the FRET signal. Subsequent demethylation separates...

... aaRSs in these processes. To create the antibody toolbox, they screened antibody candidates using a synthetic...fourteen aminoacyl tRNA synthetases (aaRS) have been created, characterized, and validated by the Gräslund lab...cross-reactivity, antibody candidates underwent affinity screening to select for slow off-rate binders. The best ...pathways By Gabrielle Clouse Claudia Vickers’ lab has created a novel nanocompartment for transporting proteins...

...detectors for all fluorophores. This allows for the creation of much larger panels, as fluorophores that cannot...they don’t “wash out” the rest of the panel and create spreading error. Conversely, markers with low expression...should not be paired with these dyes, as this would create difficulty when trying to resolve those populations...signature upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical...spectral flow cytometry has the ability to generate incredible amounts of data from a single experiment. Although...

... to its final destination. As an example, most secreted proteins are produced in the endoplasmid reticulum...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ... they are finally degraded (2, 3). Once you've created your fusion protein, it's important to validate... to make any fluorescent protein constructs you create in your own work available to the Addgene community...Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka...

...transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with a SmaI or XmaI restriction...duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary. To save time...mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial pellet...how iodixanol purifications works, shows how to create the iodixanol gradient layers, and has some great...master iodixanol AAV purification. If you’re new to creating gradients, practice making the iodixanol layers...

...suppression of background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion...obtained if any single component was omitted. Screening for successful Golden Gate Assembly All Golden..., Table 1). These improvements (5- to 12-fold increases in the number of transformants while maintaining...to faster assembly workflows that require less screening to confirm the accuracy of assembly. The fidelity...test the limits of DNA assembly approaches with increasingly complex experimental conditions. The ability...

...needs to be compensated out of APC-Cy7. After increasing the compensation values for APC against APC-Cy7...to be shifted towards the x-axis, so you would increase the compensation value for APC. If the population... to be shifted away from the x-axis, you would decrease the compensation value.   Figure 1: Compensation...population needs to be shifted towards an axis, increase the compensation value, whereas if it the population...population needs to be shifted away from an axis, decrease the compensation value. If neither of these helps...