...property by including ccdB on its destination plasmids. In the destination plasmid, the ccdB gene is flanked... interest by taking advantage of bacterial characteristics such as prototrophy, the ability to synthetize...sites. To introduce a gene of interest into the destination plasmid, recombination sites must first be added...

...mitigating noise generated by off target digestion. After MNase digestion, fragments are released from nuclei... DNA around the target protein. This targeted digestion is controlled by the addition of calcium, which...measure of DNA binding sites in combination with existing ChIP-seq data sets, of which there are many readily...

... some special characteristics of scientists in the work place as described by Sebastiano Massaro in his...some of these common pitfalls of management and still not get the most from your team. It is pretty clear...learned to get results with the carrot, and not the stick.     This is the first in a 5 part series on Introduction...

...are on the right track if they start asking you questions. Remember that people like a good story and science... this case you – on a quest to ask and answer questions and hopefully discover something new. Science ...out to science writers in your area or at your institution; the news or communications office is a good ...internship at a place like Science News. Ask your institutional news office if there might be opportunities ...

...Makarova et al. grouped the systems into three distinct types and subtypes using polythetic classification... not translatable to the other CRISPR subtypes. Still, the large and dynamic range of CRISPR systems between...viral anti-CRISPR genes. These data highlight an interesting "arms race" between phages and prokaryotes; the... and further develop the CRISPR/CAS toolkit by posting generic lab protocols, providing tips from experts...

...14] Hepatocytes [15] Pancreatic β-cells [16] Intestinal tissue [17] Lung epithelial cells [18] Red ...approaches differ in several ways. Below is a table listing some of critical differences (adapted from Zhou...Zhou and Melton, 2008, [43]): Characteristic Reprogramming+ Transdifferentiation in vivo or in vitro...differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature, 2011. 470(7332): p. ...37. Merrell, A.J. and B.Z. Stanger, Adult cell plasticity in vivo: de-differentiation and transdifferentiation...

...tips for scientists working at home, and you will still find this useful. Now that we’ve been doing this...just need to know you well enough to ask good questions and hold you accountable for your goals.  I’ve...interviews are 1:1 discussions with someone who has interesting experiences to share. The conversation builds...edit everything you’ve tried to draft, but you’ll still be practicing and getting familiar with the process...

...2016 Addgene’s research team has been actively investigating ways to improve viral vector production. Whether...Whether redesigning viral packaging plasmids, investigating different viral production strategies or developing...were unable to deduce the mechanism responsible we still believe that these results will be beneficial to...method to improve AAV yields. Although we only investigated 3 serotypes it’s possible that this approach...

...will bind to both and you will not be able to distinguish between your two targets.    Figure 1:...primary antibodies and the targets will be indistinguishable (top panel). If one of the antibodies is converted...be converted? Isotype conversion is a unique characteristic of recombinant antibodies. Recombinant antibodies... that this blog post has answered all of your questions about the utility of isotype conversions and chimeric...

...I didn’t truly address the question of what I wanted to do when I grew up until I was all but finished...writing lab reports and empirical essays had also stirred some of my more creative faculties into motion,...skill acquisition. Classes covered filmmaking, podcasting, philosophy of science, creative writing, designing...communication graduates are often well-equipped. Many institutions are sincerely interested in connecting with ...

...University, and the Karolinska Institute developed mNeonGreen via 21 substitutions to LanYFP and the addition...Nathan Shaner and colleagues from The Scintillon Institute, Allele Biotechnology, Florida State University...applications from traditional imaging to FRET and stochastic single-molecule super resolution imaging. mNeonGreen...

... have the potential to provide improvements to existing technologies based on polymerase processivity....stable, highly soluble, and non-immunogenic yet are still just as specific to their target as regular antibodies...podcast segment Steven Carr’s lab at the Broad Institute focuses on proteomics: developing and applying...analysed. In this work, the authors specifically investigated protein binding at the TERT gene, but this method...

... replicate the host's chromosomal DNA, but they still rely on the host machinery to make additional copies...Patrick says researchers can ask themselves a few questions to get started: Will the plasmid be used exclusively...that plasmids with low to medium copy numbers can still express massive amounts of protein given the proper...cells have not started to die off. Have any questions? Need something clarified? If so please, post ...

...how to amplify recombination-prone plasmids and testing your prep can save you the headache of troubleshooting...colony you selected contains the full plasmid. A diagnostic digest is one of the easiest and clearest ways...you can try re-streaking for single colonies and testing multiple preps. In some cases, we suggest gel extraction...plasmid is present, but the recombined plasmid is still visible. This indicates that the DNA prep contains...

... mountain – with many routes up and only one destination – a career in the sciences is more like an intergalactic...developmental programs unfold, absorbing the stochasticity of molecular interactions and embedding individual... weird and rare organisms so I found it hard to stick to vertebrates alone.  And a longstanding inclination...with things like word choice, punctuation, and stylistic consistency. If you’ve ever doubted the importance...

... of the primer assist with restriction enzyme digestion (usually 3-6bp) Restriction Site: Your chosen...and then TAAGCA to improve restriction enzyme digestion. This gives us a sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA... You can generate the reverse-complement using existing software (a quick internet search will lead you... The PCR product is now ready for restriction digestion. As such, the later steps in this process are ...

...labs. We’re overjoyed to provide you with these fantastic educational resources and will be sure to keep...us at [email protected]. Thank you again for a fantastic year and happy reading! Top 10 blog posts of 2015...UNC Chapel Hill explain some of the molecular gymnastics behind DNA repair by non-homologous end joining...career coaches do and offers up some career coach suggestions. 8. A Match Made in Heaven: CRISPR/Cas9 and ...

...Serendipitous Stain. https://hardydiagnostics.com/wp-content/uploads/2016/05/Hans-Christian-Gram.pdf Smith, A. ...infected lung tissue, a Danish biologist named Christian Gram realized that certain stains were retained...Stain. https://my.clevelandclinic.org/health/diagnostics/22612-gram-stain Hardy, J. Gram’s Serendipitous...

...for sleep. At Addgene, our mindfulness group is sticking with their Headspace practice even while working...for bench scientists to assist in labs that are testing for COVID-19. You can also give back by supporting...interview with you, or look for professional societies hosting virtual networking events. Download Addgene’s Science...expectations for your productivity Set a routine and stick to it. Plan defined work hours as well as time for...

... T helper type 2 (Th2) cells from mice that constitutively express Cas9 and screen for genes that affect... Steven Farber and colleagues at the Carnegie Institute for Science have developed the LipoGlo system ...UAS). The Wang Lab created a tripartite system consisting of dCas9, a gRNA fused to an RNA binding motif...displays high dynamic range. However, the promoter in question must be σ54-dependent and the system may not be...