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...this later). Chemigenetic sensors take a hybrid approach and utilize both a genetic component as well as...sensing to signal output slightly slower than other approaches. Functionally, the voltage sensitivity of the...level in vitro. In vivo monitoring has also been achieved with these sensors, but further optimization is...cell level as they are in vitro. Factors like photobleaching, lack of red-shifted indicators, and brightness...

... when I first became a manager. Know your people Each person is unique and should be managed accordingly...writing a grant, thinking about the next project, reaching out to other teams, expanding your network…. I...problem, I was able to resolve this issue - with coaching - before it became a major problem. But I also...always the case or you may not feel comfortable reaching out to your manager. This is when having a mentor...

...the genome. Native Cas9 has two catalytic domains, each of which cleaves one strand of DNA upon binding ...proteins derived from different bacterial species, each of which interacts with a distinct gRNA that allows...expression with truncated gRNAs An alternative approach is to modulate Cas9’s nuclease activity by modifying...engineering parameters via a library-on-library approach." Nature methods 12.9 (2015): 823-826. PubMed ...

...described below (See Figure 1).   Figure 1: Each problem with dimerizing fluorescent proteins is ...increasing the probability that two FPs will collide with each other. The consequences can be dramatic. For a membrane...the Endoplasmic Reticulum and live cell imaging approaches. Erik is also an avid long distance runner, gardner...14581454. PubMed Central PMCID: PMC2173526. 2. Zacharias, D.A., et al., Partitioning of lipid-modified ...

... is valuable in many experimental settings. To achieve this, scientists use a multitude of techniques ...promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic vectors simultaneously... that only a single translation event occurs for each mRNA. However, some bicistronic vectors take advantage... some of these peptides, no consensus has been reached on which peptide works best. Likely the choice ...

...it is also a good idea to run an uncut sample of each plasmid to help with troubleshooting if your digests...approximately 1:3. Since the number of base pairs for each varies, it is difficult to calculate this based ...alone. One method is to conduct 2 ligations for each plasmid you are trying to create, with varying ratios...

...transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early genes encode proteins...noting that it is possible to design a vector that achieves persistent transgene expression in certain cell...10.1080/22221751.2019.1690953. PMID: 31749409. Harrach, B. (2014). Adenoviruses: General Features. In ... Bian, A., Zhi, Q. X., Iparraguirre, A., Lopez-Camacho, C., Wherry, E. J., & Ertl, H. C. J. (2007). Adenoviral...

...neurons and GFP (green) to label neurons transduced by each serotype. Working at Addgene has shown me this... AAV vectors from Addgene and the CLOVER Center, each one submitted by researchers like you. Reports are...only be effective if many scientists choose to help each other by sharing their data. So take advantage of...214–225. https://doi.org/10.1089/hgtb.2019.105 Rumachik, N. G., Malaker, S. A., Poweleit, N., Maynard,...

...B-D represent the pure DNA spectra for comparison. Each of these contaminants will have a slightly different...that you use the same solution for blanking your machine!  Finally, many NanoDrop systems now provide full-spectrum...are no substitute for sound techniques, routine machine maintenance, and critical interpretation of the...are, of course, more considerations and other approaches for measuring DNA quantity and quality than we...

... primers. Find Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized... your plasmid Have to make a new primer pair for each insert Backbone-Specific Provides information...cloning You have to make a new primer pair for each insert since primers are insert dependent   ...

... screening), or arrayed on a 96 well plate where each well contains a defined prey fusion.  The former...possible to screen libraries of bait and prey against each other at the same time in order to try and access...2547163. 5. Ozawa, T., Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase as an optical...: PMC2965261. 8. Obrdlik, P., El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, ...

...recombinase have been developed. A common means to achieve ligand control is to fuse a mutant estrogen receptor...be transfected by in utero electroporation. To achieve sparse labeling of individual neurons one could...the same time from two different plasmids this approach becomes problematic as it reduces the probability... the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to DNA via a Phosphotyrosyl Linkage...

...ligase cereblon (CRBN), including popular new approaches like PROTAC targeted protein degraders. But CRBN...images of single focal planes through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet... abscessus either simultaneously or stepwise, achieving mutation efficiencies of up to about 9% of colonies...simplicity and efficiency of this dual-plasmid approach makes it a valuable resource for advancing our...

...are not general amplification protocols. Rather, each protocol is optimized for a single, specific library...step also depends on the individual library. So, on each library’s page, you can find protocols for sequencing...protocol on Lentivirus Production provides a general approach for producing lentivirus in Lenti-X 293T cells...virus–the right amount of virus, that is. Ideally, each cell will be transduced by no more than one viral...

...required to develop new projects and test novel approaches, a necessary skill set for any scientist. In ...lab   Thinking Outside the Box: Individual Approaches Whether it’s finding the source of contamination...your situation.   Thinking Outside the Box: Team Approaches At some point or another, you’ll find yourself...benefits from their policy! Perfectionism When we reach for perfection, our work stalls. We become so fixated...

...and organizations, like the Allen Institute, to achieve greater outcomes together. The Allen Institute... well as the strength and density of labeling in each. This allows users to quickly find the tools that...Daigle, T. L., Sedeño-Cortés, A. E., Walker, M., Kalmbach, B., Lenz, G. H., Morin, E., Nguyen, T. N., Garren...Y., Ding, Y., Somasundaram, S., Miller, J. A., Kalmbach, B. E., Radaelli, C., Gore, B. B., Weed, N., Omstead...

...process relies on “sequencing-by-synthesis,” where each base is recognized and recorded as a DNA strand ...3). It uses (aptly named) “nanopores” that are attached to the flow cell membrane and have an electrical...pulled through the pore via motor proteins. When each nucleotide passes through the nanopore, the electrical...bioinformatic analysis. At Addgene, we use a mix-and-match approach for sequencing methods and choose the best method...

... work my lab wanted to do already seemed to know each other and what to do.  Saved by the Tweet - The...people attending large meetings. A key was to provide each bin with a mentor who had attended the meeting multiple...networking and mentoring circles We looked at a lot of approaches scientific meetings have taken to provide mentoring...Joe taught high school chemistry for a year. The teaching/mentoring bug never went away. While he has frequent...

...using standard methods that have been optimized for each specific virus in order to generate high quality... generally two to three fold higher than those achieved using the standard qPCR method.  Titering by the...The specific QC experiments performed varies for each viral lot. To learn which specific QC experiments... throughout production, ensuring the identity of each sample from the DNA tube all the way to the viral...

...are many types of electronic lab notebooks (ELNs), each with its own pros and cons. All ELNs have the virtue... take a look at the RSpace ELN.  The connected approach in action: how the RSpace ELN supports reproducibility...research, and data repositories. Below I describe how each of these kinds of connectivity facilitates reproducibility...electronic lab notebook. Rory is based in Cambridge, Massachusetts, and feels privileged to be part of the vibrant...