...interest. Select for cells of interest (positive or negative selection). Next-generation sequencing (NGS) to...high degree. This is particularly helpful when investigating tissue-specific diseases, as you can target ...protein altogether. Platforming AAV Interest in interrogating and mapping the genetic landscape of tissue-...used to map tumor suppressors in mouse liver, interrogate immune cells to improve immunotherapy in glioblastoma...

...-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to...To test the library, they conducted genome-wide negative selection screens with both A375 (melanoma) and...similarly to the Brunello knock-out library while mitigating toxicity induced by high copy number dsDNA cutting...libraries. These unique hits may have been false negatives in the other screen, implying that ORF and CRISPRa...

...to spend their days microinjecting fluorescent conjugates into cells. Fortunately, over the last 20 years...Stains – These labels consist of a fluorophores conjugated to molecular species with high affinity for endogenous...fluorophore attachment, the brightness of the resulting conjugate, and the complexity of the system. A particularly...refinements to these systems—smaller tags, brighter conjugates, higher fluorogenicity, photoactivation etc.—...

...norepinephrine to promote vigilance. A recent study investigated the injection of AAV8 particles expressing an....   A study from 2014 (Howerton at al. 2014) investigated the connection between neuroinflammation and... expression of miR124a, a regulatory RNA that negatively regulates BDNF levels. They found that downregulating...prefrontal cortex (mPFC). In 2019, the Duman lab investigated this further and showed that the activity of...

...in a range of biological studies, including investigations into environmental pollutants and health, embryo...developmental biology that arose from zebrafish-based investigations. Genetic resources for zebrafish A common ...about the genome are powerful resources for investigating any biological process. High quality genome ...are particularly well-suited for neuroscience investigations. For example, neuronal activity can be tracked...

...months, in fact. In the Lee-Parsons lab, we are investigating how the production of cancer therapeutic drugs...expression method enables us to quickly screen and investigate transcription factor candidates and the regulation...transient methods are usually the first choice for investigating gene function, as these experiments can be done...different ideas. We are a huge fan of the Golden Gate based MoClo system. The vectors mentioned in this...

...pieces of DNA whose sole purpose is to ensure the propagation of themselves and their associated genes. These...specific 700 bp segment of DNA, the plasmid was propagated better than plasmids without this DNA. They eventually...Perhaps the most widely used application of CcdB is GatewayTM cloning, where the ccdB gene serves as a counterselectable...Thomas, and Kenn Gerdes. "Mechanism of post-segregational killing by the hok/sok system of plasmid R1:...

...machinery within the cell. Most plasmids encode a negative regulation system involving antisense RNAs or ...).  In cases of compatible plasmids, different negative regulation systems control plasmid replication...plasmids have the same origin of replication, the negative regulation system cannot distinguish between the... the different plasmids. The concentration of negative regulation system elements in the cell is artificially...

...The 3’ end of the non-invasive strand is elongated and ligated to fill the gap, thus completing SDSA.   ...-independent pathway as it involves the direct ligation of sticky or blunt ends and often introduces small...four-stranded branched structures that form when elongation of the invasive strand “captures” and synthesizes...

...for cloning into the final translated products. Negative selection markers built into all plasmids in the...Gagoski used SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression...for product integrity, expression level, and aggregation propensity in four expression systems: E. coli...base RNA sequence. The segments are provided in Gateway compatible vectors that allow you to quickly clone...

...adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required since...from these overhangs and ligate these blunt ends together in a standard ligation reaction resulting in a...

...innovative tools enabling large-scale real-time interrogation and monitoring of brain cells and circuits during...

...success. Gather data on the lab choices How do you find out what the lab is really like? Gather data of... lab website, publication records, LinkedIn, ResearchGate or Twitter. Be ready for the interview with ...Alumni meetings can be the most informative way of gathering data. Former lab members can give open, revealing...

...repressible promoters can be regulated via positive or negative control. Positive Repressible: An activator protein...sequence anymore and transcription is turned OFF. Negative Repressible: A repressor protein is present, but... as sphingomonads and bacillus.  ADH1 The ADH1 negative repressible promoter is commonly used in yeast...that promoter is active. In this way, you can interrogate the activity of uncharacterized promoters. Similarly...

...can be particularly useful for sharing negative data. Negative data often goes overlooked, but knowing... not encoding this protein, which we will call Negative Control Gene (NCG) (A). Then you lyse the cells...antibodies, as well as a third unrelated antibody as a negative control (A). The two antibodies to your protein...

...account name in the top right corner of PubMed to navigate to your account settings. Next, click on the link...Preferences” box. Once on the Preferences page, navigate to “PubMed Preferences” and click on the “Filters... deposited plasmids. Now that you know how to navigate to the PubMed Preferences and Filters pages, be...

...is, of course, interesting to parse through, aggregation of this information can provide valuable insight... started to explore how I could visualize the aggregated data. Plotting the data in interactive figures...Since Addgene encourages the public sharing of aggregated data only, information on requesting PI(s) & ...

...love negative data, and in fact, what we're most hoping to see from the community is your negative data...the data when an AAVs doesn't work. Sharing your negative data helps others to design better experiments...

... of starting material; Gibson assembly or Golden gate assembly use a more direct approach, allowing scientists... a lipopolysaccharide of the cell wall of Gram negative bacteria like E. coli that affects tissue culture...differences between transformation, transduction, conjugation, and transfection Resources on Addgene.org ...

... just above it can help you quickly and easily navigate to educational resources, tools, collections in...options into a single place, so you can quickly navigate around the Addgene website without scrolling up...disabilities will find Addgene’s website easier to navigate, thanks to design choices informed by Web Content...