...experiments. MET15 L-methionine and overproduces hydrosulfide ions yes - Grow with methylmercury. S. cerevisiae...to high copy numbers. This creates a potential metabolic burden on the yeast cells. Some phenotypes may...tried varied approaches to combat these issues. One method to reduce the amount of marker gene expression ... utilizing the more traditional drug selection methods feasible in yeast as a complement or alternative...

...swamped out by a sea of uncoupled donors. The best method for targeting a fluorescent donor to your protein...tRNA synthetases is required and thus far, this method has proven most effective only in a few cell types...This is not an extensive list Acceptor labeling: Sometimes it's okay to be wrong In contrast to FRET donors... (which was originally developed to treat heavy metal intoxication in chemical warfare [Vilensky & Redman... of your FRET assay (Figure 2). These labeling methods are freely interchangeable and new uses for FRET... structure using a novel site-specific labeling method." PloS one 4.10 (2009): e7338. PubMed PMID: 19823671...dynamics in living cells with FRET reporters." Methods 40.3 (2006): 279-286. PubMed PMID: 16908183. 10...

...’s science communication journey started with something unexpected: termites. “I was really interested... work for one client and then switch to doing something else. You’re never really integrated and part ...pulled in all of these different directions is sometimes at odds with the space needed to be creative. ...a lot of time to let you mind process things. Sometimes finding that uninterrupted time to be creative...some articles are just harder to write. “There’s something about being a writer that can be a very painful...these problems through connections and that’s something that good storytellers in science can do.” Susan...

... a visual aid instead of describing something verbally. Sometimes a group was just really tired and needed...as it does to read an 11-page Cell paper - and sometimes, it might actually take me less time to read the...concepts are easier to grasp than others. But sometimes we’re unaware that we are leaning on an unspoken...way I thought about engaging with learners. If something I was communicating didn’t click, instead of blaming...put their phones away), or maybe I was assuming something was intuitive that actually wasn't. Maybe I needed...

...What about all the reagents you need to order? Sometimes, it feels like an eternity of cloning, waiting...costs of producing plasmids with high-throughput methods that save on reagents, consumables, and time. We...first try (or any response at all). And while this method is free for you, and many labs are quite happy ...strategies beyond the traditional PCR/digest/ligate methods and picking the right one can save you time if ...speed or money if budget is a concern. The faster methods, like Gibson assembly, are typically more expensive... 5 factors to help you choose the right cloning method Find blog posts about scientific sharing here Download...

...you’re probably aware that there are several methodologies currently available for approaching it. These... traditional restriction enzyme/ligase-mediated method, the more recently developed Gibson Assembly Cloning...cloning technologies, as well as several others. Each method is unique and relies on specific components that...components is essential for choosing the correct cloning method for your own experiments, and here we will focus... a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB, what role does... to ensure that you choose the most appropriate method for your own experiments. Please check out Addgene...

...largest family of transcription factors found in metazoans. The superfamily is comprised of 48 human and ...a broad range of reproductive, developmental, metabolic, and immune response programs. NRs also encompass...gallstone disease, senescent illnesses, and the metabolic syndrome. Coregulators Coregulators constitute... regulatory regime (such as genes involved in metabolism, growth or cytokine action). Primary or core ...and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature afforded...adipogenesis and macrophage activation. Our mass spectrometry-affinity purification proteomic profiling of...

...interrogate neuronal networks. A major drawback of this method is the stray excitation of neighboring neurons ...material for the bioproduction of therapeutic methylxanthine drugs like theophylline. To facilitate future...enables the degradation of caffeine (1,3,7-trimethylxanthine) and its conversion into xanthine. The majority...constitutive promoter (BBa_J23100) and expresses N-demethylases and other required enzymes, including a glutathione...rescued by the xanthine produced via complete demethylation of caffeine. For example, the Barrick lab was...Find the FLINC biosensors here. Mo et al. Nat Methods. 2017. PubMed PMID: 28288122. New fluorescent...

... to some of your potential options and cloning methods, but please scroll down for plasmids suitable for...inserted using Gibson Assembly or SLIC cloning methods. A BsaI-based E. coli multiplexing plasmid is available...multiplexed gRNAs with Golden Gate and Gibson assembly methods If you want to scale up the number of gRNAs in ...your plasmid, you’ll need to use some assembly methods such as Golden Gate or Gibson assembly. Golden ...containing destination vector. Note: depending on which method you use, the procedure will vary slightly. ...Fok1-dCas9 (dimeric nuclease). Gateway assembly method Frew Lab Multiple Lentiviral Expression Systems...Plasmids: The CRMAGE system is a fast, multiplexable method that combines CRISPR and recombineering-based MAGE...

...You have to weigh the pros and cons of various methods and choose what is best for you — and the mice....potential pain or discomfort of the mice, and restraint method. Many different substances can be injected, including...on the back. Depending on the site and restraint method, subcutaneous injections take some practice to ...injections is studying inflammatory responses, as this method is useful for analyzing localized events. Intradermal...the throat. The feeding tube can be flexible or metal, depending on researcher preferences. It sounds ...injection routes in mice! When considering what method you’d like to use, remember that the final decision...

... production or looking to improve your current methods, understanding the differences can help you make...will depend on your specific virus, so this is something you may have to optimize for your specific application...the virus type. AAV is typically purified using methods like ultracentrifugation, while lentivirus supernatants...up to 38 kb! Generally, insect cell production methods produce lower titers of virus than mammalian systems...systems. This is why many insect production methods add an additional viral passage. This involves taking...taking your first round of virus, called P1 (or sometimes P0), infecting new Sf9 cells, and getting an amplified...

...of interest and examine them using biochemical methods thus removing the context of the cell. However,...important to choose the appropriate quantification methods and tools. It is usually not enough to examine ...review of quantitative colocalization analysis methods and useful image analysis software tools. Pitfalls...Pitfalls, limitations, and specialized colocalization methods Fluorescent proteins are very useful tools, but...further investigation. One fluorescent microscopy method commonly employed for analyzing interactions between...fluorescent protein for cellular imaging." Nature Methods 14.1 (2017): 53-56. PubMed PMID: 27869816. 6. Media...

...ice, whichever is more practical. If your lysing method is not resulting in your protein being available...need such precise measurements, you can use a colorimetric ladder, visible to the naked eye; a fluorescent... of protein, make sure your reversible staining method of choice is appropriately sensitive. If using ...membrane, you can dry the membrane, wet it with 20% methanol, and place over a light box to visualize proteins...that the p membrane pores are too large.  Transfer method   Wet transfers are more efficient, so if you’re...re-run the western.  Remember that chemilumescent methods, like horseradish peroxidase protein (HRP), have...the bubbles in the membrane sandwich Ensure methanol concentration in transfer buffer is < 20% ...

...AAVs for neuroscience research is the lack of a method for genetically manipulating neurons throughout... Plasmids Here! Their Solution: Use the CREATE method to engineer two new IV injectable AAV capsids, ...for studying the CNS and PNS! What is the CREATE method?  CREATE stands for Cre Recombinase-based AAV Targeted...axons and dendrites extend to. The keys to this method are 1) high levels of color diversity so that cells...system is less cumbersome than brainbow, another method for multi-color labeling of neurons, because it...capsids (figure 1) were engineered with the CREATE method (figure 2) to allow for efficient IV delivery of...

...are in the process of transitioning our titering method from qPCR to ddPCR. Droplet digital PCR (ddPCR)...your PCR For a good starting place on what PCR parameters to use, see Lock’s seminal paper on AAV titration...between positive and negative droplets with these parameters, here are a few things you can try: Increase...in faecal samples." Journal of microbiological methods 148 (2018): 64-73. PubMed PMID: 29548643. Lock,...titers by droplet digital PCR." Human gene therapy methods 25.2 (2013): 115-125. PubMed PMID: 24328707. PubMed...Kristina, et al. "A systematic investigation of parameters influencing droplet rain in the Listeria monocytogenes...

...and its related fluorescent proteins, which are sometimes used as tags for detection; however, those are...and cellular resources are being spent to make something the cell doesn’t need to make. Eukaryotes and ...function, His-tagged proteins can be purified using metal-affinity chromatography, usually using a Ni2+ column... used for western blots, IP, co-IP, IF, flow -cytometry; can occassionally interfere with protein folding...HBH HHHHHHAGKA GEGEIPAPLA GTVSKILVKE GDTVKAGQTV LVLEAMKMET EINAPTDGKV EKVLVKERDA VQGGQGLIKI GVHHHHHH  9 ... used for western blots, IP, co-IP, IF, flow -cytometry, but rarely used for purification as elution requires...

... work and the rest of life is sometimes more of a struggle and sometimes less. When we are lucky, we are... make a change. #2 Thou shalt sometimes put work ahead of life Sometimes, work has to come first. That...responsibilities are less You can work at home sometimes, but do so carefully and with attention to those...ready for anyone in the family to have to miss something – for example, say no to some of those birthday...

...genotyping, and biotechnological advancements. Many methods of nucleic acid detection however, either lack ...of the Cas13 protein to develop and optimize the method termed Specific High Sensitivity Enzymatic Reporter...s non-specific ssDNA degradation to develop the method termed DNA Endonuclease Targeted CRISPR Trans Reporter...(SNPs) from human saliva. SHERLOCKv2 provides a method to detect nucleic acids with high sensitivity and...compromising speed, ease of use, and portability. This method can be used for an array of applications including...Myhrvold C, Freije CA, Gootenberg JS, Abudayyeh OO, Metsky HC, Durbin AF, Kellner MJ, Tan AL, Paul LM, Parham...

...Sanchez and Alice Ting. Sanchez MI et al., Nat methods. 2019. https://doi.org/10.1038/s41592-019-0665-...localization of biomolecules within cells. However, sometimes these fluorescent proteins might not localize ...doi.org/10.1101/160374 Nanobodies- there's a simple method for that! By Michelle Cronin The benefits of nanobodies...Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion body problem and...could be easily purified by one-step immobilized metal ion adsorption chromatography (IMAC) using Ni-NTA... human cells) is a CRISPR-Cas13 based antiviral method to degrade RNA from SARS-CoV-2 sequences and live...

...networks that regulate processes from development to metabolism to fertility. Several zebrafish-specific gene...Cas9. Another popular targeted gene deactivation method in zebrafish are short, modified oligonucleotides... generating mutants. Unexpectedly, phenotypes sometimes differ depending on whether a gene is deactivated... approaches (Harland, 2018). Several different methods have been developed, relying on established reagents...application for Cre/lox recombination experiments." Methods in cell biology. Vol. 135. Academic Press, 2016...resolution in freely swimming zebrafish." Nature methods 14.11 (2017): 1107. PubMed PMID: 28892088. Nüsslein-Volhard...