... to determine the pros and cons of using many different types of gRNA design software. When you are happy...yourself through whatever DNA supply company you currently use. You can also find many previously validated...inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a...weeks old and use the ear tag method (creating different patterns of ear holes and using that material ...latest and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target...plasmid into zygotes." Development, growth & differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541...

...sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different RTKs. Although...Improving on the glutamate sensor iGluSnFR from Loren Looger’s lab, Kaspar Podgorski and colleagues have...response. They generated variants optimized for different applications (v82 has higher affinity for glutamate...By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato...sub-millisecond resolution and sub-degree temperature differences under a variety of imaging conditions (155 μs...

... iPSCs (or correct them back to wild type), differentiate them into somatic cells affected by the disease...iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones! We ... experiments with blocking mutations, we still weren’t entirely happy with our editing outcomes and in...Dominik Paquet and Dylan Kwart Dominik Paquet is currently a Professor for Neurobiology at the Institute ...Tessier-Lavigne’s lab at the Rockefeller University. He currently works to understand pathological mechanisms of...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering...

...labs. While many labs approach the situation differently, some adopting simple note-taking tools, others...functionalities with experiment records. Many different types of ELNs are available on the market today...available here. Our questions focused on researchers’ current practices, their opinions towards ELNs, and their...desired functionalities i.e. main priorities of different ELN features, ranging from whether respondents...cases this applies to universities. How are you currently exchanging information with your team? If you ... allows you to assign specific permissions to different lab members to avoid any issues later on. Do you...

...tagged protein because of its size. This type of interference is known as steric hindrance. For instance, ...OSER whorls. This assay is quantitative and is currently the standard for assessing oligomerization in ...fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus. For...advisable to generate multiple constructs with different insertions, since it is difficult to predict which...can follow him on twitter: @joachimgoedhart.   References Adjobo-Hermans, Merel JW, et al. "Real-time visualization...291.5512 (2001): 2408-2411. PubMed PMID: 11264536. Klarenbeek, Jeffrey, et al. "Fourth-generation epac-based...

...similarities with humans. However, rats have some key differences in physiology, intelligence, and human correspondence... generate transgenic axolotls for an array of different studies. Elly Tanaka’s lab has constructed several...include understanding of allelic variation between different plant species and metabolic properties affecting... collections to help start your experiments! References: Blount, Z. D. (2015). The unexhausted potential...Zhang, C., & Xing, X.-H. (2018). Pooled CRISPR interference screening enables genome-scale functional genomics...Plasmids Collection Page Read our molecular biology reference Find plasmids for your research ...

...solubility, and thus different reagents are useful for biotinylation in different microenvironments. There...used today in molecular biology for an array of different techniques and protocols. In this post we will...determined to be due to a deficiency in Vitamin H later renamed Vitamin B7/biotin. Egg white injury would plague...a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular...post-translational modification of exceptional specificity.” Trends in Biochemical Sciences, 24.9 (1999b): 359–363....

...scientists have developed and fine tuned many different ways to clone DNA fragments which have provided...with an enzyme master mix, which contains three different enzymes: an exonuclease, which chews back the... this doesn’t allow for mix and match of many different fragments. One way around this is to use a combination...with a G!) but actually work by significantly different methods. Golden Gate cloning does result in the...from one vector to another. There are lots of different ways to clone these days. Gibson and the other...biology, you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC,...

...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the monkey story hit the presses, another very different bit of CRISPR news was circulating. Scientists...how CRISPR/Cas9s actually work. Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales...this structure may help us engineer around the current limitations of the Cas9 complex," said study author...

...Toronto, created The Addictive Brain to spread awareness about how scientists utilize tax-payer money and... how our articles have helped them understand different topics of science, especially disease related ...terms like manipulations, anomaly, and so on, differently than scientists would. For example, “manipulate...blogger, Chinmaya Sandangi! Dr. Chinmaya Sadangi is currently a postdoc at the University of Toronto and founder... receive our science career guide Learn about different careers in science communication Resources on...

...fragments that are very different.” Because the fragments produced will have different overhangs, they can...systems. “The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann...Lohmann of Heidelberg University. “What is different is the plasmids you use to bring this into your target...available, making it easier for us to do our work.” References 1. Athanasios Lampropoulos et al. "GreenGate ...

...5,000 different research institutions. This has given us a unique window into technology trends in the...by the influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids...HA-Ubiquitin-WT and HA-Ubiquitin, and we now have 19 different ubiquitin plasmids in the repository. 100,000 ... and modified by so many labs and for so many different purposes that we had to create a CRISPR page to...

...by Hannah Dotson in Dec. 2022.  References and Resources References Gossen M, Bujard H. Tight control...In the presence of tetracycline (Tet), TetR preferentially binds Tet over the TetO elements, allowing ... One can therefore place GAL4 and GAL80 under different promoters and create sophisticated patterns of...wondering, “why is it necessary to have all 3 of the different binary systems above if they all behave very similarly...tet-on-and-tet-off-2nd-generation Mullick, A., Xu, Y., Warren, R. et al. The cumate gene-switch: a system for...

...line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin,...rearrangement (such as when a stem cell initiates differentiation), transgene silencing may occur (as it would...cassette can be incorporated, and used during the differentiation process. Moreover, a splinkerette-PCR assay...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-...piggyBac-ing models and new therapeutic strategies.,” Trends Biotechnol., vol. 33, no. 9, pp. 525–33, Sep. 2015...26299960. PubMed Central PMCID: PMC4559351. 8. A. G. Uren, H. Mikkers, J. Kool, L. van der Weyden, A. H. Lund...

...controlling properties like growth, survival, and differentiation.” https://www.addgene.org/kits/sabatini-wood-cancer-pathway-orfs...implicated in cancer cell proliferation, survival, differentiation, and apoptosis. For each of these 17 pathways...morphological features, migration, or survival in different contexts.” It’s also possible you could make use... sometimes drug screens and sometimes totally different experiments we never thought of - and that’s thrilling...constructs to the kit in the next two years. References 1. Martz, Colin A., et al. "Systematic identification...

...tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community. Don E...PiggyBac transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered...is a new member of this class that works via a different mechanism - the PhoCl protein is irreversibly ...“caged” by fusion to specific domains thereby rendering them inactive in the cytoplasm. The authors use...interest. NHEJ- or HDR-induced mutations in ATP1A1 can render cells resistant to the small molecule ouabain -...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ...examining genomes possessing CRISPR systems with apparent self-targeting. Two of these Lmo proteins, AcrIIA2...

... Conversations with children playing in the background became a normal occurrence as Addgene and our university...light of the need for basic research during the current pandemic, universities are showing an unprecedented...of Illinois Chicago                     Boston Children's Hospital                           California...scientists. This partnership complemented each other's strengths: Ginkgo Bioworks’ infrastructure allowed them ...

...neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists... populations using viral strategies Combining different approaches can increase the specificity of cell...optogenetic payload in neurons that project to different regions of the brain. A more locally defined set...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...share experimental design and data. With these different approaches to open science, we aim to accelerate...

...year bioengineering student, to Addgene. I am currently working on the QC/QA team as a DNA Sequencing ... food service industry, which followed a very different schedule. Fortunately, I was provided with a detailed...gave me enough time to become familiar with the different types of issues that were important to this function...Specifically as a co-op, it is encouraged to explore different teams and work on projects there. At this point...point, I have seen several lab processes and am currently working on projects for the Science Communication...

...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three colors. The...specificity of these orthologs is key: due to differences in the PAM sequences required by each ortholog...specific for telomeric sequences and showed that different fluorescently labeled dCas9 isoforms are efficiently...target sequence. They were also able to label two different pairs of chromosomes using gRNAs specific to sequences...Addgene are excited to see what’s coming next! References Multicolor CRISPR labeling of chromosomal loci...

...trained to be good at learning new things—this is a strength you can draw on and demonstrate in your application...be excited to be there. If you are appying for diffrent types of positions (for example, bench research...resume should also be customized to highlight different skills and to get the important information in...hiring manager. Learn to confidently describe your strengths in your resume and don't be shy about asking for..., you will never get the help that can make a difference. Having your resume forwarded personally gives...

...antibodies, and more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, ...Nonetheless, this tag can be purified with several different resin types and is a fairly small tag that is ...  Affinity tags vs. other tags There are many different types of tags, including epitope tags (FLAG, Myc..., D. W., & Wingfield, P. T. (Eds.). (2001). Current Protocols in Protein Science. John Wiley & Sons, Inc...

.... Northeastern compiles a list of hundreds of different jobs for students to apply to, and after a grueling...marketing and social media co-op. There are a lot of different worries that come along with entering the workforce...and co-founder, approached me and asked about my current projects and personal interests. We chatted about...participated in the discussions, I began to note trends in the data, and from that, suggest changes to ...of my co-op. I had to do research on loads of different best practices, create an email strategy, and ...

... tissue, in vivo. Two of their constructs are currently available as Cre-dependent, EF1ɑ-driven viral ...have high selectivity for K+ over Na+ and rapid photocurrent kinetics. They demonstrate the use of HcKCR1...two-photon excitation around 1,040 nm, has larger photocurrents, and is more selective for K+ than HcKCR2, making... be used as a potent optogenetic silencer. The current work also proposes promising avenues for future...anti-c-Myc [9E10] (isotype IgG2a) antibody is currently recommended for use in western blot, but we are...

...and insecticidal proteins to the development of renewable biotechnology products and biosensors for herbicides...Development of a bacterial expression system for renewable biotechnology products Dr. Idah Sithole-Niang,...database for this important agricultural plant. Her current research focuses on endophytes, bacteria, and fungi...effect observed in bacteria. Learn more about the different CRISPR applications in plants.     Transgenic ...helped us coordinate and promote the awards. References 1. Martinez-Klimova, Elena et al. “Endophytes...

... Substrate: Some luciferase reporters, such as Renilla luciferase (RLuc) or NanoLuc, require injection...together genes from gas vesicle operons from two different bacteria, Bourdeau et al created two constructs... 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles is the amount ... invisible to ultrasound, which allows for differentiation of arg1 and arg2 ultrasound signals. This also...and arg2 plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan...

...leader-specific. How is leadership different from good management? Aren't all career skills leaderhsip skills...because it emphasizes the importance of these very different roles. At any stage in our careers we may be in...you are not fulfilling the requirements of the current task. Most of us start as doers, some become managers...boss) that you are ready to try operating in a different role?  I think the secret sauce can be summed ...

...Cong worked with Addgene Senior Scientist, Matt Ferenc, to organize the CRISPR forum's most popular questions...double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will... like the wildtype Cas9 does. There are a few references for the double nickase system, including one ... Sanger sequencing). Click here for a useful reference.  More FAQs, CRISPR protocols, and gRNA design... and optimized for specific plasmids Links to different software to help you identify your gRNA target...

...the issue is not unique to IHC.  References and Resources References Goodpaster T, Randolph-Habecker J...endogenous IgGs in the tissue. Created with biorender.com. Strategies for avoiding background secondary...them to your sample so the secondary antibodies aren’t free to bind endogenous IgGs. (C) A third solution...primary and secondary antibodies. Created with biorender.com. Proper tissue processing and sample preparation...

...this emerging technology! Resources and References References Orguc, S., Sands, J., Sahasrabudhe, A., ...electrophysiological potential afterwards. This measurement trended with expression of the opsin in the target neurons...obvious next applications are to expand the tool to different brain-organ axes. Alternatively, the system could...Free Behavior. 2020 42nd Annual International Conference of the IEEE Engineering in Medicine & Biology...