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We narrowed to 850 results for: met

Showing: 281 - 300 of 850 results

  1. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...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...

  2. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...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...

  3. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...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...

  4. Plasmids 101: Protein tags

    Type
    Blog Post
    ...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...

  5. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ... 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...

  6. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...mix-and-match approach for sequencing methods and choose the best method based on the plasmid. By working ...sequencing deviates from the synthesis-based sequencing methods (Figure 3). It uses (aptly named) “nanopores” that...hour to 72 hours, with real-time data output. This method is great for resolving complex and repetitive sequences...their processes, quality, and efficiency through methods like automation. All assemblies from our pipeline... has also improved efficiency in many of their methods to pass materials through the QC process quickly...

  7. Hot Plasmids - August 2020

    Type
    Blog Post
    ...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...

  8. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...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...

  9. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...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...

  10. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...the only method of determining drug resistance status, it is a very common, well-trusted method that is...of acquired drug resistance are referred to as methods of ‘horizontal gene transfer’. Resistance can also...Drug resistance is not a black or white issue. Sometimes bacteria are resistant to lower concentrations... will be killed by a higher concentration, or sometimes more exposure time is needed for the drug to work...of inhibition around the disk is measured in millimeters, and corresponds to the level of susceptibility...

  11. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...weeks, but, in this post, we will outline a simple method for selecting the guide RNA, validating its efficacy... first. In vitro screening is a simple and fast method for validating your selected gRNA’s, and has the...RNAs encoding Cas9 into the oocyte. This original method was developed as the use of CRISPR/Cas9 in mammalian...deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a finger,... the pups are 2-4 weeks old and use the ear tag method (creating different patterns of ear holes and using...material for genotyping), but bear in mind that this method will require you to house mice that may have no...

  12. The Twelve Days of CRISPR

    Type
    Blog Post
    ... to me: SHERLOCK and DETECTR, two CRISPR-based methods to detect nucleic acids.  On the third day of CRISPR... fifth day of CRISPR, Addgene gave to me: five methods for detecting SARS-CoV-2 using CRISPR.  On the ...seventh day of CRISPR, Addgene gave to me: CRISPR methods for bacteria.  On the eighth day of CRISPR, Addgene... the tenth day of CRISPR, Addgene gave to me: a method to capture promoter-enhancer 3D chromatin structures...the eleventh day of CRISPR, Addgene gave to me: methods for adenoviral delivery of CRISPR/Cas9.  On the...

  13. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...neuronal differentiation. Nat Methods 14:963–966 . https://doi.org/10.1038/nmeth.4430 Nihongaki Y, Kawano ...Gene editing Manna et al., 2019   NPPOC-caged trimethoprim (TMP) Cas9 fused to destabilizing domains (Cas9...proteins for optogenetic control of CRISPR–Cas9. Nat Methods 15:924–927 . https://doi.org/10.1038/s41592-018....201606123  Kim JH, Rege M, Valeri J, Dunagin MC, Metzger A, Titus KR, Gilgenast TG, Gong W, Beagan JA, Raj...looping for endogenous gene expression control. Nat Methods 16:633–639 . https://doi.org/10.1038/s41592-019... CRISPR: A New Relationship Built to Last. In: Methods in Molecular Biology. Springer US, pp 261–281 Manna...

  14. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...oldie but a goodie — a phrase used to refer to something that has fallen out of fashion but is still useful...technologies have gradually taken over as the preferred method for many labs. However, RNA interference (RNAi)...quickly rose to popularity. Compared to CRISPR, RNAi methods are often quicker and the repositories of validated...synthetic constructs introduced through external methods. shRNA processing Following delivery into cells...this is a common issue with many gene silencing methods and just needs to be taken into account when designing...Design, delivery, and assessment of gene knockdown. Methods in Molecular Biology, 139–156. https://doi.org/...

  15. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...with this, it’s a method for building and testing new products or methods, using metrics and feedback, taking... videos for free. We’re a small operation, so sometimes it can be a big challenge. But ask any filmmaker...for longer than anticipated, we had to think of something else.   Figure 2: Ironing boards. Good ...need as much detail as possible when showing lab methods. So, this just wasn’t going to work for the majority...

  16. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...or gel filtration is the most common analytical method. However, these may not reflect the situation in...results.  The recent revolution in genome editing methods (most notably CRISPR/Cas mediated HDR) enables ...Quantitative assessment of fluorescent proteins." Nature methods 13.7 (2016): 557. PubMed PMID: 27240257. PubMed...characterization of dedicated sensors for FLIM, for ratiometry and with high affinity." PloS one 10.4 (2015)...community-editable fluorescent protein database." Nature methods (2019): 1. PubMed PMID: 30886412. Mastop, Marieke...monomeric orange and red fluorescent proteins." Nature methods 5.6 (2008): 545. PubMed PMID: 18454154. PubMed ...

  17. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ... this system to be used to trace neurons over millimeter-long distances across multiple fields of view...instead of XFP to optimize it for tissue clearing, a method that makes tissues, such as the brain, transparent...Chemical tags are more resistant to tissue clearing methods and can be labeled by synthetic fluorophores, which...Improved tools for the Brainbow toolbox." Nature methods 10.6 (2013): 540. PubMed PMID: 23817127. PubMed...circuit analysis in Drosophila melanogaster." Nature methods 8.3 (2011): 260. PubMed PMID: 21297619. Köhler,...homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells." Cell 143.1 (2010)...

  18. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control...Spinach-based riboswitches can be used as stable metabolite sensors for imaging in live cells. By placing...placing Spinach downstream of a riboswitch, metabolite binding to the riboswitch instead activates Spinach ...Spinach fluorescence. In the search for something more stable for quantitative imaging, Ming Hammond’s group...and U6) and measuring Corn fluorescence by flow cytometry, the Jaffrey group was able to quantify the effect...of trinucleotide repeat–containing RNA." Nature methods 10.12 (2013): 1219. PubMed PMID: 24162923. PubMed...

  19. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...stably or transiently depending on the delivery method and cell type in question. CRISPR components can...wall has been removed). Another common delivery method is agrobacterium-mediated delivery, which uses ...In contrast to homology directed repair, these methods do not involve double strand break (DSB) formation...wheat by Caixia Gao’s lab. More recently developed methods can also be used to efficiently deliver CRISPR-...used, precise protein sequences or domains, and methods of delivery - the underlying technique of CRISPR...crop plants using the CRISPR/Cas system. Plant Methods 9:39 . https://doi.org/10.1186/1746-4811-9-39 Gao...

  20. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...behave very similarly?” The simple answer is that sometimes it is necessary or desirable to control or look...expression and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed...expression system in Caenorhabditis elegans.” Nat Methods 9(4) (2012): 391-5. PubMed PMID: 22406855. PubMed...Q-system reagents for genetic manipulations.” Nat Methods 12(3) (2015): 219-222. PubMed PMID: 25581800. PubMed... A Versatile Expression System for Drosophila. Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939...Srikrishnan. “Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae...
Showing: 281 - 300 of 850 results