...Nobody’s perfect! This is a platitude I’m sure we have all heard. It’s a truth that extends to both people... of tools that rely on DNA amplification and manipulation to produce and edit plasmids. These are prone...polymerases can make mistakes and lead to the accumulation of errors in DNA sequences. In research, these...and identifying mixed plasmid and nucleotide populations. All of these topics require additional considerations...sequence and assemble using the Illumina MiSeq platform, such as GC-rich regions and highly repetitive...Plasmidsaurus for long-read nanopore sequencing. This platform sequences the entire plasmid in one read and thus...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in over...expression and pseudotyping efficiency. This plasmid was later used by another team of researchers for neutralization...opportunities to evolve. Several variants are now circulating and plasmids containing these mutations have ...are inhibitors of SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV...their work, head over to their blog post. Months later, we partnered with Ginkgo Bioworks to provide synthesized...screens identify mucins as critical host factors modulating SARS-CoV-2 infection. bioRxiv. https://doi.org...

...field as it provides a noninvasive method for manipulating cell activity, especially neuronal firing. ...approved drugs so that your results are viable for translation to human patients. Picking receptors The receptor...dependent on what cellular response you aim to manipulate. It is important to consider that the same receptor... in. The most common target for chemogenetic manipulation are neurons where chemogenetic receptors can...control ion flux. The system allows you to directly manipulate ion channels. When working with PSAM receptors...BL, Jin J (2015) The First Structure–Activity Relationship Studies for Designer Receptors Exclusively Activated...

...calls, I feel compelled to share my years of accumulated wisdom so that you, the customer, can get the...these tips and tricks will apply to any biology-related customer support interaction. Before contacting...paperwork associated with a product only to realize later that everything I could have wanted to know was ... science, but we don’t have the resources to recapitulate the experiments performed by our thousands of...use a particular reagent.  We also have links to later papers that successfully used said plasmids. We ...support scientists wear many hats at this still relatively small nonprofit. If you do go longer than a few...

...City The conference was located in the modern, relatively new “Science City” area on the outskirts of Guangzhouon...human tRNA genes and, until now, we've overlooked regulatory fragments of these transcripts. I remember when...have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs. Stay tuned...Medicine presented another unexpected finding in the regulatory RNA field. The Rajewsky lab is studying circular...of Georgia studies the CRISPR/Cas system as it relates to its natural role in prokaryotic immunity to ...against both DNA and RNA viruses - 2 cas systems regulate at the DNA level and 1 at the RNA level. Udi Qimron...

...other systems because the system modulates protein post-translationally, unlike many other methods. Secondly... tags has become increasingly popular for the modulation of endogenously and exogenously expressed proteins...systems. Degron tags are one of many tools to manipulate protein expression. These tags offer several ...Secondly, these tags can be used to manipulate endogenously and exogenously expressed proteins. Thirdly...degradation. Since its first use as a protein regulation tool, AID has gone through an additional round...

...any antibody (positive population) or won’t bind anything (negative population). It is strongly recommended...immunologists, allowing for the rapid analysis of cell populations within a heterogenous tissue type, such as PBMCs...moreso than other methods, the identification and isolation of rare cell types. In recent years, full spectrum...for better resolution of positive and negative populations, Careful panel design In both conventional and...create difficulty when trying to resolve those populations. PE/Cy7 and BB515, on the other hand, have a ...

...DNA in your sample relative to your normal control - you can set up your calculations so that your assay... one-copy ‘housekeeping’ gene), and quantified relative to a 'normal' sample with known copy number set...nice thing about qPCR is that since it measures relative quantification - that is, since you’re looking...all double-stranded DNA, while Taqman uses an intercalating dye, with a fluorescent probe and quencher to...provides absolute quantitative data, instead of relative quantification.  Even though no one option is ...

...important to understand that these platforms, or pieces of a platform, that are now available exist within...well-characterized bacterial transcription and translation initiation elements. Open science with BioBricks...replication of DNA, transcription of DNA to make RNA, translation of RNA to make proteins. That's central to everything... transduce otherwise invisible molecules into regulation of gene expression including growth or fluorescence...gene expression via standard transcription and translation initiation." Nature Methods. 10, 354-360 (10 ...

...about how they became grant professionals, mid- to late-career grant professionals have similar stories ...’s requirements as well as state and federal regulations. A grant professional’s job duties can range ...close-out a grant. Grant professionals also build relationships with foundations and government grantmakers....keep up-to-date on the changing guidelines and regulations impacting grant management, we track grant opportunity...amount of grant funds available and the increasing regulatory process for grants has fostered the demand for...worked for a health department and then for the regulatory services department as our healthy homes program...

...However, retroviral vectors can still direct the translation of proteins because they integrate this reverse-transcribed...then use cell machinery for transcription and translation. Here, we will discuss several aspects of the...The genome of viral tools are often not virology related at all, and you will instead get expression of ... Cullen, B. R. (1992). Mechanism of action of regulatory proteins encoded by complex retroviruses. Microbiological... sarcoma virus: purification of the virus and isolation of the nucleic acid. Proceedings of the National...Initiation of synthesis with 70 S viral RNA as template. Journal of Molecular Biology, 79(1), 163–183....

...more focused format, where all students work on malate dehydrogenase (MDH), a citric acid cycle enzyme...recently purified MDH to study the impact of phosphorylation on the enzyme’s structure, function, and interaction...offering excellent training in improvisation and the latter making him a more effective motivator (“be like...conducting NIH- and NSF-funded research on planarian flatworms both in and out of the classroom.   References...characterization of a fusion protein: Histidine tag, malate dehydrogenase, and green fluorescent protein. Biochemistry...laboratory – a semester-long research approach using malate dehydrogenase as a research model. Biochemistry...

...vector - overall, I spent two years testing and reformulating our AAV construct to optimize performance in...Working at Addgene has shown me this was far from an isolated case. Our scientific support team answers hundreds... AAV questions every year on a range of issues related to AAV vectors, many from people trying to answer.... (2020). Methods Matter: Standard Production Platforms for Recombinant AAV Produce Chemically and Functionally...its occurrence during vector purification and formulation. Molecular Therapy, 12(1), 171–178. https://doi.org...Researcher Perspectives on Data Sharing in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December...

... science.                    Two of the platforms we use at Addgene: YouTube, for longer educational...octogenarians is going to be a hard sell!) Digital platforms often have demographic data of their users readily...geographic location. For example, not all social media platforms are available or used in all countries; not all...the limitations and potential of your preferred platform before you start can save a lot of trouble in ...requirements.  No visuals. Variety of hosting platforms can splinter audience. Primarily digital distribution...communicators. Well-developed content creation platforms.  Algorithms can make it difficult to reach people...

...GFP derivatives, but are instead related to the dsRed protein isolated from Discosoma sp. Similar work ...common fluorescent proteins and their mutations relative to wildtype GFP. Although not listed here, many... fluorophores Fluorescent Protein Mutations Relative to Wildtype GFP EGFP F64L; S65T EYFP S65G;...that separates mixtures of cells into distinct populations based on fluorescent signal. Thus, FACS can be...has also been used to identify particular cell populations in drug screens, to visualize micrometastases...

...receiving the translation, Addgene contracted with a professional scientific translator, L. Denise Roche...101 eBook is now available in Spanish!  This translation was gifted to Addgene by Daniel Mejías Sánchez...Roche, to edit the translation. We are pleased to announce that CRISPR 101: Una herramienta de edición ...  Many thanks to:  Daniel Mejías Sánchez, PhD, translator  Denise Roche, PhD, MD, editor, ldeniseroche@...

...the ratio of genome-containing viral particles relative to the total number of viral capsids (which can...vectors consist of full particles (white arrowhead) relative to empty particles (black arrowhead). Scale bar...measured by qPCR.   Physical titer is typically calculated by two popular PCR-based methods: Quantitative...is a factor in AAV titration since the starting template is the purified viral vector solution (which contains...be mitigated by using purified viral DNA as the template instead of the intact AAV particle. Finally, ...absolute titer values may not be as important as the relative titer values. Finally, because physical titer ...

...L-arabinose) without signs of toxicity or burden relative to a fluorescent protein control. We also added.../L yeast extract, and 5 g/L NaCl) or on LB-agar plates containing glucose and L-arabinose at 37oC for ... arabinose and colonies on arabinose-containing plates will be visibly white, and gas vesicles in cells...from left to right: colony whiteness on inducer plates, white inclusions in cells with GVs via phase contrast...heterogeneity required single cell screening to isolate a clonal cell line capable of producing ultrasound...homogeneity, as the cells could be batch-sorted to isolate a polyclonal cell line with strong gas vesicle ...

...space — developing tools for cellular targeting. Lately, they’ve been working on two tools that expand ...safe to say the sample had their attention! They isolated DNA from the sample, amplified the CRISPR locus...most common RNA targeting system, Cas13, the "collateral" activity of the enzyme can lead to off targets...this information to identify a large variety of related bacteria with similar Cas systems. Some of these...knockdown efficiencies. The ability to identify related Cas systems opens up the potential of further development... also compatible with both AAV and adenovirus templates. The system, which is larger than Cas9, can also...

...Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether with...affinity, were eventually called aptamers, from the Latin aptus, meaning “to fit”. SELEX could be used to ... protein fluorophores and quenchers are still relatively bulky. One of the advantages of oligonucleotides... context of a paired fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their...hydroxybenzylidene imidazolinone (DFHBI) with relatively low fluorescence in vivo, the Jaffrey lab used...