...centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013. These terminators can...: 2388-2393. Huang, Shijie, Xuechen Zhu, and Charles E. Melançon III. "Detection and Quantification...metalloproteinase: López-Pelegrín, Mar, et al. "A novel mechanism of latency in matrix metalloproteinases." Journal...size from 4 to 156 kDa, enabling the user to characterise the efficiency of their cell-free system in ...5373-5383. Walkiewicz, Katarzyna, et al. "Small changes in enzyme function can lead to surprisingly large...

...designed to be a helpful desktop reference. Each chapter brings together all of our 101 blog posts on an...to antibodies as research tools, you may find “Chapter 1: What is an Antibody” quite useful. If you’re... design your first flow cytometry experiment, “Chapter 7: Sorting” is there to help guide you through ...find an antibody that works, we highly recommend “Chapter 3: Finding the Right Antibody for Your Experiment...

...Cre-dependent conditional alleles, has remained a challenge. Enter Easi-CRISPR: a method that harnesses the...First, you’d construct a plasmid with your desired changes, long homology arms and a selection marker, then...template. Although effective, ivTRT may be more challenging for sequences with complex secondary structure...-purified DNA template.   Many thanks to Channabasavaiah B. Gurumurthy, Masato Ohtsuka, and Rolen M....PMC5434640 Miura, Hiromi, Rolen M. Quadros, Channabasavaiah B Gurumurthy, and Masato Ohtsuka. “Easi-CRISPR...

...positively charged groups, such as aldehydes in fixed samples. To address this, block positively charged groups...the label control for all new samples, protocol changes, or when high background labeling occurs (Burry...critical for data interpretation, especially when changes in protein expression are the experimental readout...       References and resources References Burry, Richard W. “Controls for Immunocytochemistry.” Journal ...

...promoter, and titer.     Figure 1: Flow chart depicting a) steps for selecting serotypes and promoters... different receptors, co-receptors, and other mechanisms to gain entry into the cell (Dudek et al., 2018... different brain regions.  Image modified from Aschauer et al., 2013, Figure 1. Used under Creative Commons...the region you want to cover - and as your titer changes, so may your needed volume (de Backer et al., 2010....2014.00076 References and Resources References Aschauer, D. F., Kreuz, S., & Rumpel, S. (2013). Analysis...

...added, it binds tTA, resulting in a confirmation change that prevents binding to the promoter and turning...added, it binds cTA, resulting in a confirmation change that prevents binding to the promoter and turning...this way, you can interrogate the activity of uncharacterized promoters. Similarly, placing UAS upstream ...1472-6750-6-43Kaczmarczyk A, Vorholt JA, Francez-Charlot A. Cumate-inducible gene expression system for ...genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197...

...had record readership with over 75,000 views. Changing Labor Laws Bring Increased Postdoc Wages Our most...information for anyone considering a postdoc. In “Changing Labor Laws Bring Increased Postdoc Wages,” Future...for some of these institutions to manage these changes quickly and harmlessly. Learn much more about the...Reasons to Use Reddit for Science Communication Changing Labor Laws Bring Increased Postdoc Wages I hope...

...to face global challenges, we hope to empower those that need to face these challenges locally. We look...themselves on the front lines of emerging global challenges. However, these same researchers face significant...international distribution As you can see from the chart below, we are making great progress toward spreading...

...Antibodies are composed of two light chains and two heavy chains. The ends of the branches contain the...variable, allowing exquisite specificity. This characteristic allows monoclonal antibodies to be used as ...the DSHB, play an important role in scientific exchange. In addition to ensuring affordable access to ...

...the RNAi trigger delivering the RNAi trigger characterizing the RNAi phenotype. Although all these steps...plants to have enough tissue for downstream characterization. To simultaneously address both limitations...the RNAi output and trivializes downstream characterization, as the entire culture can be harvested instead...Stainier DYR (2020) Transcriptional adaptation: a mechanism underlying genetic robustness. Development 147....186452 Wilson RC, Doudna JA (2013) Molecular Mechanisms of RNA Interference. Annu Rev Biophys 42:217–...

...kinase activity is FRET, but FRET reporters are challenging to design and difficult to multiplex. The Covert...which fluorophores can be multiplexed together. Challenging  to Design: FRET reporter activity depends on...and NES sequences, which have previously been characterized. See Box 1 in Kudo et al. for more details on...Resources on the Addgene Blog Learn about photoswitchable kinases Learn more about fluorescent biosensors...

...Act on information to make change whenever possible. Be public about changes influenced by feedback. If... Human Dynamics Laboratory identified five characteristics of very successful teams. Check out Joanne'... post in this series). Members carry on back-channel or side conversations within the team. We are ...

...the lines between science and art was Albert G. Richards, who passed away in 2008. His work exposes and... use of x-rays. His haunting images have been enchanting people for years and have gathered much acclaim...Smithsonian Magazine, where the editor stated “Albert G. Richards, a teacher of dental radiography, had focused ...which are now extinct. A collection of Albert G. Richards’ work is held by The Museum of Jurassic Technology...

.... “When PhD students graduate there has to be champagne, that's a given.” Furthermore, anyone leaving ...fellowships, jobs, or paper submissions with dinner and champagne in the lab. The Church Lab celebrates successful...toasts a newly minted PhD with food, cake, and champagne. What does your lab do for fun as a group? The...those have been trips to breweries who happily exchange our expertise with lots of free beer.” He also...

...application process can dramatically increase the chances of being seriously considered for an open position...steps to submitting a resume that will give you a chance at an interview:  1) Have an excellent resume ready...or location. The more people you meet the better chance you have of finding a beneficial connection. LinkedIn...resume forwarded personally gives you a much better chance at an interview –but you still need to nail the...

...covalent bonds, so there were no weak links in the chain.  Addgene: What about the Streptococcus pyogenes...antibody tentacles may have an impact on this challenge either for magnetic capture or for capture using...this tool? Howarth: Polyproteins are useful for mechanical study of proteins by atomic force microscopy ...particular tips or tricks for those wishing to purchase and use the SpyLigase system for the first time...

...appropriate antibiotic. Although the precise mechanistic details of the process are not well understood...resultant products of the reaction in the figure from Chaokun et al., 2011 to the right. After PCR, you should...with any PCR-based cloning technique there is a chance of introducing an unwanted mutation. Tips for high...(2016): 145-154. PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free...

... to students.” Making science approachable is often the most challenging part of the job, but Caitlin ...lab classes. However, their opinions are often changed after seeing a kit for themselves. “You open up...Center. And make sure to subscribe to the Career Channel of the blog so you don’t miss the future posts ...

...are ever recycled, so adopting a few meaningful changes in the lab could make a big difference in our global...Many scientists are already leading the cultural change toward responsibility in our lab’s environmental...Labconscious. If you’re inspired by this post then I challenge you to take to action! These days, it’s important...’s up to all of us to work on this issue. Be a champion for this important cause! To get more lab sustainability...

...40 cas gene families have been characterized. To date, 83% of archaeal and 45% of bacterial genomes utilize...defense against invading phages. Rising to the challenge - Evolution of anti-CRISPR genes in phages The...Makarova K, Haft DH, Barrangou R, Brouns SJJ, Charpentier E, Horvath P, Moineu S, Mojica FJM, Wolf YI, ...