... plasmids in the repository, each verified by sequencing, which makes the repository a convenient source...content and rare codon percentage were not good predictors. There is not much variation in GC content between...for GC content. Rare codons also are not good predictors because they are just that: rare. “Nature has...says. Percent sequence identity was a different story - the team found that the percent sequence identity... a synthetic version of that gene was a clear predictor of whether the gene is synthetic or natural. They...most common expression system within the Addgene repository is mammalian, but the largest source of unique...is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed that the...

...Topoisomerase I is released from the vector when the PCR product and vector ligate; however, it can potentially...this sequence. The energy from this breakage is stored as a covalent bond between the cleaved 3′ DNA strand... days commercially available TOPO kits provide vectors or cloning arms with overhanging 3´ deoxythymidine...residues that are covalently linked to topoisomerase. Vectors in these kits also often have the topoisomerase...screening after transformation - self joining of the vector ends results in the production of blue colonies...Reaction: Mix together the PCR product and TOPO Vector.  3. Incubate 5 Minutes at Room Temperature: You...are planning to transform right away OR you can store the reaction at -20C overnight. 4. Transform TOPO...

...molecule integrin inhibitors and natural ligands, such as fibronectin, to block receptor function, e.g.,...technologies and laboratory techniques related to antibody discovery. You can find cell surface receptor biology...Integrins are essential cell surface receptors involved in complex signaling pathways and the linkage...Despite their ubiquity and significance, these receptors have proved challenging to study. Cells often ...integrin antibodies available through Addgene’s repository. The collection includes 26 unique antibodies...from its closed to its open state, activating the receptor. Ligands can bind to integrins in their closed...subunit. Using these affinity reagents, the investigators inhibited other RGD-binding integrins, individually...

...characteristics that make them ideal for studying in a laboratory environment. We’ve covered five model organisms...are five more model organisms that have left a historical impact on the scientific community. (And, check...have been specifically developed to thrive in laboratory settings. One of its most attractive features...Level 1 microbe makes it a perfect candidate for introductory teaching and research environments (Idalia &...  The E. coli genome has an extensive research history and characterization. Many proteins and processes...studies. Elly Tanaka’s lab has constructed several vectors for use in the axolotl including plasmids for use...organisms in the world. With its extensive scientific history, highly characterized genome, and widespread utilization...

...deaminase-mediated base editing (cytosine base editor or adenine base editor) and reverse transcriptase-mediated ... et al., 2019). Both cytosine base editors and adenine base editors have been developed for plants and...CRISPR has taken the genome engineering world by storm owing to its ease of use and utility in a wide variety... al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9-VP64) or repressors (dCas9-KRAB...variety of Agrobacterium-compatible, Cas9 containing vectors ready for Golden Gate mediated cloning of your ... the Patron Lab.  As with all plasmids in the repository, we highly recommend reading the associated publications...Z, Wang Y, Wang X (2019) Rescue of a plant cytorhabdovirus as versatile expression platforms for planthopper...

...contributed by guest blogger Natalie Niemi, a postdoctoral fellow at the Morgridge Institute for Research...phosphorylation sites. If knowledge of a candidate regulatory kinase is known, it’s possible to generate non-phosphorylated...Often, however, researchers lack knowledge of the regulatory kinase, access to active kinase for the reaction...tRNA with phosphoserine A specialized elongation factor that allows proper extension and translation of...synthetase, and EF-Sep – an optimized elongation factor that accepts Sep-charged tRNAs during translation...This modification allowed the deletion of release factor 1, the protein responsible for terminating translation...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can ...

...the abundance of a protein variant library is monitored by fusing the variants to eGFP followed by an ... is an enzyme critical for activating clotting factors and is the target of the anticoagulant drug warfarin...serotonin sensor The goal of Elizabeth Unger’s postdoctoral work in Lin Tian’s lab at UC Davis is to generate...serotonin is a well-known but poorly studied neuromodulator, and evidence suggests that it is incredibly...sensor). In preliminary studies with the lab’s collaborators, they were able to use the serotonin sensor ...activity. Unger also plans to use the serotonin receptor to study how single nucleotide polymorphisms present...and dose response. Selective serotonin reuptake inhibitors are frequently used to treat depression and anxiety...

...upon a moratorium until another meeting, three branches – a private funder, government regulators, and ...there was a generally agreed-upon international moratorium on utilizing CRISPR/Cas9 in humans. This topic...mtDNA transfer options being investigated; A UK regulatory body tasked two separate non-governmental groups...technology to research centers engaged in long-term, mandatory follow-ups and observational studies of the technique...whether or not scientists can recognize so-called bad actors among them and “self-police” or whether the media...to argue otherwise). To be sure, throughout the history of medical research, there have always been people...stop punting decisions on scientific, ethical, regulatory, and general public consensus to the next meeting...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...activation, using expression of differentiation factors as the screen readout. This retroviral mouse CRISPR...library can be used for screening of cells that are refractory to lentiviral transduction, now enabling forward...lipoproteins, shuttles lipoproteins through the circulatory systems, but can also lead to the formation of...as both a powerful tool for generating complex regulatory systems and further the study of natural σ54-... PMID: 31451697. Genetically encoded voltage indicators for use in worms Article contributed by Brook...Brook Pyhtila Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when neurons...

... fields, including synthetic biology. Addgene depositors Christopher Voigt and Jeffrey Tabor have been...and inactivate biological processes. Chemical effectors are suboptimal for a number of reasons, including...cross-reactivity with other pathways. Since these effectors are diffusible, they’re also not suitable for ...histidine kinase that phosphorylates a response regulator (RR). TCS-induced responses are varied; one outcome...conditions, the chromophore activates the response regulator via phosphorylation, promoting transcription of...spread across three plasmids, with the response regulator encoded in the genome. To make the system less...The utility of these TCSs was limited by a few factors. These bulky systems were spread over a number ...