...Overall the general strategy for protein expression consists of transfecting cells with your DNA template of... Mammalian systems can express proteins both transiently or through stable cell lines. Both methods produce...recombinant proteins from plants however can be expensive and time consuming as plant tissues themselves...applications. Cell-free expression systems however, are expensive and very technically challenging to use. Alyssa...

...all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently dimerize. That means that your...to not produce artifacts in your experiments. Considerable effort has gone into making FPs monomeric, yet...actually monomeric, at least in practical terms. The propensity of a particular type of molecule to form a dimer...study of GFP-based FRET, with application to intrinsically unstructured proteins. Protein Sci, 2007. 16...

...RNA-guided gene drives in the summer of 2014 (after extensive discussions with biosecurity experts, ecologists...gene drives intended for eventual release should consider building an appropriate immunizing reversal drive...not use them whenever applicable?Against this, consider the cost of an accidental release. Science relies...trusted to handle such a powerful technology responsibly. An accidental release would be particularly ...

...DDBJ, and PDB sequences and may be the most comprehensive for searching. Timesaving Tip #1: If you know...searches! Now, before you click the BLAST button, consider the Program Selection parameter, as this will ...the alignment at a more distal amino acid. We consistently run our blastx searches with the “Low complexity...101: How to Verify Your Plasmid Plasmids 101: An Inside Look at NGS Plasmid Quality Control Learn about...

...translation. Its genome consists of “early” (E) and “late” genes, and the transition between early and late...length. Human adenovirus type 5 (hAdV-C5) can be considered the prototype, although many viruses infecting...the majority of adenoviral vector expression is transient due to the fact that AdV surface proteins (especially...

...specific genomic region using the CRISPR system, consisting of dCas9 and targeting gRNA, to identify molecular...isolation of a specific genomic region was an intensive process. One method required insertion of a recognition...vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and synthetic ...high-affinity purification. This system is both highly sensitive and specific and allowed the Xu lab to identify...

...data with a 7-day rolling mean gave us a crude insight into the magnitude of the effect COVID-19 had on...holiday effects. Based on these observations, we considered the following (noiseless) model for our data:...factor 𝜙z if t is during event z. The "events" we considered for our model were weekends, Christmas/New Years...Europe. To improve the stability of our model, we considered only the 10 papers with more than 10 unique order...

...Working with AAV vectors in neuroscience Let’s consider my first challenge when I started working with...and that it often provides incredibly valuable insight into one’s work. As a community, we must therefore...., Grames, M. S., & Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector...with Addgene’s AAV Data Hub!Data Freedom: The Expansion of Data Sharing in Research Publications...

...method into six general steps and outlined the considerations to be made during each step. Now that you know...method of fixation is good for epitopes that are sensitive to cross-linking, but it doesn’t preserve sample...binding to any target protein that is still on the inside of the cell. However, assuming you do need to permeabilize... then optimizing from there is a good option.  Consider your target - if you are studying a membrane protein...

...abandoned early on due to its considerable immunogenicity and its transient expression. LV results in long...been shown to to transduce the retina but only transiently. However new viral vectors capable of providing...disease-causing genes, including the RPE65 gene responsible for an autosomal recessive, severe, childhood-onset...

... in transit and some have been transported across oceans and continents. Each deposit may consist of anywhere...your lab ensure that sample ID’s are unique, consistently labeled, and that numbers are not switched around...Resources Avaliable on the Addgene Blog Read more Inside Addgene Posts! Learn all about Plasmids in our ...

...research grants, especially in the sciences and by extension to scientifically oriented nonprofits. Like Deb...career path changed when I was asked if I would consider consulting for a particular grant by another state...entering the Grants Profession. Whether you’re transitioning from a previous career or you’re starting your...above and formalizes your development. You may consider earning a degree in Grant Writing, Management ...

...these challenges, researchers have amassed a considerable molecular tool box of tags and fusion proteins...deploy proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical...EINAPTDGKV EKVLVKERDA VQGGQGLIKI GVHHHHHH  9 Combo Consists of a bacterially derived in-vivo biotinylation...and Gly residues   This article is not a comprehensive guide to all tags, but rather a quick overview...

...letters, while higher-level jobs are more likely to consider cover letters an important part of the application... content team, where writing is a core skill, considers cover letters to be very important. On the other...unofficial poll, hiring managers also said that they considered cover letters to be more important for higher-level...communication skills” try giving an example, like “I have extensive communication experience, having given more than...

...that evolved in the last 10,000 years (which is considered recent). They have diverse traits with good genetic... and one is promiscuous and the gene that is responsible is understood. It’s possible to actually silence... to get a recombinant because this process is expensive. They are using a cool fluorescent indicator system...trying to replace embryo injection (hard to do, expensive, not every lab has the capability) with receptor...

...included tests done in cell lines known to be more sensitive to toxicity from RNA editing, such as mouse embryonic...CRISPR-driven precision into a genome, can be considered the fourth generation of CRISPR, one which drops...drops itself neatly into that gap. Consider, for instance, a disease like cystic fibrosis, which can be...that this system wasn’t dependent on the cell’s intrinsic machinery being already activated to work. (It...

...The ACE reporter is a bicistronic construct that consists of a mutated mCherry and a downstream constitutively...that results in the restoration of fluorescence. Transient reporter for editing enrichment (TREE) The Wang...Wang and Brafman lab developed a transient reporter for editing enrichment (TREE) to purify single base...Schwarz G, Nguyen T, Wang X, Brafman DA (2019) A transient reporter for editing enrichment (TREE) in human...

...systems that are widely present in bacteria. They consist of a toxin which can affect a variety of cellular...division”). The encoded toxin-antitoxin system consists of CcdB (toxin) and CcdA (antitoxin) where CcdB...systems Because restriction modification systems consist of two enzymes that have opposing activities, it...the systems just described, the hok/sok systems consist of three components: Host killing (hok), a long-living...

...show a high density of cells, while contour lines that are spread apart show a low density of cells (Figure... a ‘gate’; it tells the machine that everything inside the gate is one population and everything outside...Heat maps are often used to indicate the relative density of cells. You can see both of these features in...

...within Cas13a are essential for RNA cleavage, consistent with known roles for HEPN domains in other proteins...located at the 3’ end of the spacer sequence and consists of a single A, U, or C base pair. In bacteria,...bacterial cells will likely be limited by the propensity of Cas13a to bind and cleave RNA non-specifically...Finally (and perhaps most interesting), Cas13a’s propensity to cleave any RNAs after binding a user-defined...