...screen (Koveal et al. 2022). Lactate is a key metabolite in glycolysis, so this sensor will be a valuable...valuable tool to study cellular physiology and metabolism under various conditions. LiLac has improved response...going viral! In addition to the viral preps noted above for iGluSnFR3 and JEDI-2P, several biosensors already...innovation next year! Did we miss any? Let us know about your favorite new biosensor plasmids in the comments...

... all desired codon changes (step 1 in the image above). A second universal primer is used to synthesize...first followed by the top strand. Both produced about the same number of positive clones. See Supplementary... of DNA degraded in the first digest (step 1 in above diagram). Remember that the BbvCI enzymes nick the...mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How would you use...

... one of the MEK1 phosphomimetic residues in the above study with phosphoserine (to create a pSer/phosphomimetic...non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid - Addgene...interested in the intersection of cell signaling, metabolism, and the mighty mitochondrion. You can follow...Blog Verify Your Constructs with Colony PCR Read about Tags for Protein Purification Resources on Addgene.org...

...compile more comprehensive demographic information about the postdoc work force, problems start with the ...by Paula Stephan is $40,000) Increase Knowledge about Postdoc and Graduate Student Career Outcomes University...through many iterations read by your PI, your collaborators, others you know in the field, and even people...important questions that still need to be answered about the makeup and welfare of the many researchers out...

...integration of multiple copies of the construct (about 8 in Brainbow-1.0 mice.) The combinatorial effect...with the goal of overcoming the limitations listed above. First, they screened a variety of fluorescent proteins...method has become very useful in lineage tracing; “Zebrabow” was used to trace the development of the corneal...: 10.1038/nmeth.2450. Epub 2013 May 5. PubMed. Zebrabow: multispectral cell labeling for cell tracing ...

...is an off-target effect. Should I be concerned about off-target effects? Depending on your experimental...this could be a huge concern, or nothing to worry about all. Let’s consider several scenarios below for ...Predicting off-target events As mentioned in the above section, there are gRNA softwares available which...diligence to ensure they are indeed a non-issue. Above we mentioned how to predict off-target frequency...

... (post or pre run in the table above, respectively). Some of the above stains require you to visualize...equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid alone...get experimenting!   Additional Resources: Read about Gel Stains on the BitesizeBio Blog  Additional ...

... “transitivity,” calls into question any claims about specificity of a given RNAi technique. While transitivity...conceptually similar to the microRNA precursor mentioned above and is a popular RNAi trigger in the mammalian research...substantial ways: variable silencing efficiency and labor-intensive processes to isolate RNAi plants. With...resource on the Addgene blog Browse our blog posts about plant biology Check out our CRISPR blog posts Resources...

...mycoplasma contamination in your cell culture or you are about to begin a new cell culture project. If mycoplasmas...that are difficult to trace. These include the laboratory personnel, the serum, the cell culture media,...accounts for the largest source among those mentioned above. The contaminants can spread through dirty clothing...proliferating shortly. Once they are present in the laboratory environment, they are difficult to eradicate....

...et al. 2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...unbiased sequencing may not be worth the cost and labor when single clones are selected. Conversely, translational... Hough is a Biochemistry PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute...

... infrastructure, and secure collaboration.  In that post, we talked about Addgene’s data processing needs...Last summer, we posted about our bioinformatics quest, which led us to Seqera Labs, a company that provides...

...call Epi-fluorescence microscopes. (See left image above for an example image). Point Scanning Confocal Microscopy... plane to reach the detector and all light from above or below is excluded. Parallelized Confocal Microscopy...due to limitations in light penetration mentioned above (5). Thick static samples Ex: Fixed tissue sections...sectioning. Point scanning confocal microscopes (see above) often provide the highest quality images for this...

...fluorescence microscope where the light source is mounted above (epi) the specimen and the excitation light passes...technique that allows imaging of living tissue up to about one millimeter in depth. Two low-energy photons ...response, but the study of these dynamics was still laborious as epifluorescence microscopy wasn’t able to give...skin. Two-photon intravital microscopy. The video above depicts the infiltration of neutrophils expressing...

...community.  You'll also find tons of information about Addgene's ready-to-use antibodies, viral vectors...and four! So you'll also find Addgenies out and about, attending sessions, visiting posters, and leading...Sunday, from 12 pm - 2 pm. If you want to learn about, and practice, principles of clear, accessible writing...

...expedite the application process.  More information about export control licenses is available through the...Industry and Security. If you have any questions about this or other Addgene shipping and ordering processes...customer support team at [email protected].  Curious about how rabies plasmids are used in the lab? Check out...

...scientists using AAV to share practical information about their experiences. It will contain all of the juicy...data to find patterns that we can share with you about promoters, serotypes, expression patterns, and more... more. If you have feedback or suggestions about the AAV Data Hub, we welcome you to share your thoughts...

...supporters of science communication. To help you think about ways to effectively communicate your science, we... of programs SITN runs and learn how they think about communicating science. Listen to the interview ...Interview with Science Rapper Tom McFadden Learn about SITN's work at DayCon 2015 5 Reasons to Use Reddit...

...researchers time by quickly relaying key points about an unfamiliar topic or lab process. Along with our...goals, or to answer any questions you might have about Addgene. The page is divided into three main sections...a variety of roles at Addgene.  You’ll hear all about their career paths, future goals, and get some useful...

...multiplexing options, but also check out our blog post about multiplexing with Cpf1.   Figure 1: Multiplexing...assembly.  Unlike some of the plasmids described above, this vector does not contain Cas9, so you’ll need...similar to the Csy4 polycistronic system described above, except that they use an endogenous nuclease system...The Proper Cas9 Variant for Your Experiment Learn About Multiplexing with Cpf1 Additional Resources on ...

... Cepko. Listen to the podcast below to hear all about the research being conducted in Professor Cepko'...and mentoring styles. If you'd like to learn more about Professor Cepko's recent work on developing GFP-...the time to talk to us and look forward to hear about more exciting discoveries from her lab. Special ...