...us to the key question: what is the best way to compare the photostability of different fluorescent proteins...measurements for different fluorescent proteins and by comparing how the fluorescence intensity changes over time...photostability of the fluorescent proteins can be directly compared. An example of a photostability measurement of...fluorescent protein can be identified by a head-to-head comparison of several fluorescent proteins of the same spectral.... In figure 1 we provide an example of such a comparison between cyan fluorescent proteins using widefield... power, the photostability of the variants is comparable (lower panel of figure 1). To avoid changes in...

...specific chromosome. This image represents the composite of fluorescent measurements from red, blue, and...modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly, thereby decreasing the amount of background... labeling efficiency was similar to that of a comparable FISH method. Importantly, they did not observe...with the previously established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that...that they could evaluate the degree of chromatin compaction even for this small distance! To the authors’...CRISPRainbow generates 6 unique colors and a white composite. gRNAs with two identical hairpin binding domains...

...specific components that are key to the cloning reaction. Understanding the specific components is essential...take up your insert. But exactly how does ccdB accomplish this? Let’s start with a brief history of the...system contains ccdB. A successful insertion will completely replace ccdB with the investigator’s insert of...interest.   CcdB resistant E. coli strains complete the system But ccdB alone isn’t the only key to...protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3):735-45. PubMed...

... and trolls. People don’t like to connect with complete strangers as it borders on my first rule of networking...’m asked a lot about requesting connections to complete strangers for informational interviews. Many have...option is in the “More…” menu. You can also follow companies and organizations to see their news and job postings...interviews.  Pro tip: You can learn a lot from the company LinkedIn page. This can include news, inclusion...It’s also a great way to keep track of past accomplishments. Follow all the rules of good resume writing...writing. Include examples of concrete accomplishments in both the technical and “plus skills” areas. Include...

...and the efficacy of your competent cells (when they get old, they lose competency). If all the above is ... be sure that the buffers and temperature are compatible between the different enzymes. Always check the...source (i.e. primer3) or provided by the major companies selling PCR reagents. Nowadays commercial polymerases...a general rule, try to use an excess of insert compared to the backbone plasmid; a starting point could...plasmid. Many vectors contain sequences that are complementary to standard sequencing primers. If not, design...

.... Addgene is now as much a tech company as it is a reagent company. I used to have to worry about this...remarkably so for a 90 person company. We were and remain ahead of most companies in creating a welcoming, ...often focus on the excitement of what we are accomplishing and what more we might do. I am grateful every...innovative thinking, and openness to team work.   Company culture Once in awhile I worry a very little ...

...presentation, lost in a sea of data, my mind went completely blank. After several seconds of truly panicked...honest, if my paper hadn’t been ridiculously more complex than the one before it, I don’t think I would have... panicked state, only option one would let me accomplish my goal.   As a scientist, public speaking isn't...dumpster noises were just last week!  Nerves It’s completely normal to be anxious, nervous, and/or apprehensive... in its most difficult form. You’re presenting complex data, to a group of peers who are there to provide...conferences, or even just provide quick updates during a company meeting increases visibility of your work, opens...

...was contributed by guest blogger Alan Wong. The complexity of biological systems can hinder our attempts...simple tool that allowed us to rapidly decode the complexity? The CombiGEM-CRISPR technology was developed...of providing an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your ...mapping the combinatorial genetics that underlie complex regulation in biological systems. The CombiGEM ...new technologies to interrogate and understand complex biological systems. He can be contacted at aslw...transcriptional activation by an engineered CRISPR-Cas9 complex." Nature (2014). PubMed PMID: 25494202. PubMed ...

... do as genetic circuits become larger and more complex? Voigt: If you look at all the products of biology... on genes at different times. It requires more complex synthetic networks. Addgene: What hurdles did you... perspective and you can layer them for more complicated functions. Those things are easy with CRISPR..... There haven’t been enough genetic parts for complexity. We have characterized almost 600 terminators...of a cell. The objective is to give people the complete toolbox for taking on very large genetic engineering...just using what biology has given us. Wood is a complicated material and we can access it, but we are stuck...

...science and collaboration. The field already has comprehensive reference pages and curated databases for scientists... (CeNGEN) The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron...that is creating the “first virtual organism in a computer.” This project has produced various resources ...four-dimensional contrast interference (DIC) microscopy and computer imaging processing. The combination of these methods...methods provides unique opportunities to develop computational methods to better understand animal development...Hammarlund, Marc, et al. "The CeNGEN Project: The Complete Gene Expression Map of an Entire Nervous System...