We narrowed to 791 results for: REP

...up colony PCR reactions is nearly identical to preparing a standard PCR reaction: combine template, primers...other colony PCR tips are highlighted below. Preparing template: Pick a single colony with a sterile ... cultures with the clones you pick and only mini-prep the positive ones. Regardless of which method you... a few positive clones, the last step is to mini-prep these clones and submit the plasmids for Sanger ...

...primate neocortex. Cell Reports, 34(13), 108754. https://doi.org/10.1016/j.celrep.2021.108754 Panigrahi,..., STX) to the brain. Figure 2: Selection, prep, and testing of AAV enhancers in mice. Adapted from...evaluate the resulting expression of the fluorescent reporter, the brain tissue was extracted, sectioned, and...

...activity, such as Fos, Egr3, and Arc. Thus, CRISPRa represents a powerful method to dissect the individual and...astrocytes, and oligodendrocytes, provided that reporter genes specific to these cell types are used. The...including studying mRNA dosing effects, cellular reprogramming, and disease modeling. Further, the paucity ...identify factors that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed...

...expression using other methods.  In terms of preparation, VSV-G is stable and can withstand ultracentrifugation...The glycoprotein will be incorporated into the replicating rabies virus envelope, travel retrogradely, and...Lentivirus Get tips for titering your lentiviral preps Resources on Addgene.org: Check out the lentivirus...lentivirus guide Find ready-to-use lentiviral preps at Addgene ...

... are institutions doing? Institutions are now preparing to comply with the changes to the law on Dec 1st...National Postdoctoral Association Institutional Policy Report 2014). However, ~30% set their minima lower. Furthermore... 340 institutions listed as having postdocs, representing ~85% of the postdoctoral workforce. Based on... the 2015 and 2016 Future of Research Symposia Prepare Yourself for a New Career in Science with Our Career...

...share- as opposed to sell- science. 4. You can prepare answers beforehand. Joanne answered many questions...questions to provide moral support and to monitor user replies, but I can’t thank Joanne enough for all the work...incentivize good mentorship: AMA mechanics: Be prepared! The mechanics of doing the AMA were interesting...interesting. I started watching the questions and preparing answers early in the day. Although I didn’t start...

...impressive on a larger scale, too. The first paper reporting genome editing by CRISPR in mammalian cells appeared...reviewed in Lander, 2016). The first base editor was reported three years later in 2016 (Komor et al., 2016)...on one allele of his CPS1 gene, which could be repaired with an A→G edit on the complementary strand. ...enzyme requires a PAM sequence of NGG (where N represents any base), their engineered variants are more...

...Bunch of baby spinach, or BoBS, contains tandem repeats of Baby Spinach. It was developed in a study of...growth as RNA biology, we are looking forward to reports on the next additions to aptamer soup! References...fluorescent rna aptamer spinach2 for imaging of replication and immune-mediated changes in intracellular ...Spinach2 reveals the dynamic nature of trinucleotide repeat–containing RNA." Nature methods 10.12 (2013): 1219...

...finer points of your techniques. You DO need to be prepared to know the literature of your field and all of...absolute minimum, can appreciate their research. Be prepared to discuss other people's research. Nothing turns...by email or in person. If you feel compelled to repay me in some way, I encourage you to help your friends...with their job searches. If you feel the urge to repay me personally, I will gratefully accept a very hoppy...

...), and width (W). The different measures are represented as FSC-A/SSC-A, FSC-H/SSC-H, or FSC-W/SSC-W, ...describe the shape of a histogram, which graphically represents the time and intensity of the cell’s illumination...cytometer (before you set the first gate) will be represented through FSC-A vs. SSC-A, so most people use that...scatter (FSC/SSC), the detected photons can be represented through a graph of photocurrent vs. time. The...

...often used for animal germline modification, to repair or add in disease-causing mutations, but, until...suited for genome editing. The homology-directed repair (HDR) pathway is downregulated in mature tissues...Although the three DMD studies referenced here represent a great step forward for CRISPR gene therapy, ... and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit Genes Read How Non-Homologous End...

...addition, PVDF membranes can be repeatedly stripped of an antibody and reprobed with different antibodies allowing... (SDS-PAGE) is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content...control. In these cases, the western should be repeated.  Finally, while westerns are a useful way to ...

...mutation, one generally relies on homology directed repair (HDR) to incorporate new information into DNA. ...cut site is >30nt from the proximal ends of the repair template (Yang et al., 2013). This means that, ...the authors note that “clonal heterogeneity may represent a more serious obstacle to the generation of truly...efficiency of CRISPR/dCas9-mediated transcriptional repression. Nucleic Acids Res 44:e141–e141 . https://doi.org...

...infect defined neuronal subpopulations, and this represents a major downside in optogenetics (Belzung et ...gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to easily detect opsin-expressing...behaviors (de Jong et al., 2019). FLEx vectors represent, therefore, an ideal partner to optogenetics to...recombinase in mouse primary afferents." Scientific reports 8.1 (2018): 7321. PubMed PMID: 29743652. PubMed...

...subtypes, A–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first...adaptations for increased fidelity, activation/repression, high precision edits, and more. Figure...Schematic of the basic CRISPR-Cas9 mechanism, representing type II-A. Created with BioRender.com. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named A–I and...

...ligand-binding assay. Scientific Reports, 6, 25769. https://doi.org/10.1038/srep25769  Grimm, J. B., & Lavis, ...ligands for dopamine D2/D3 receptors. Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020...mitochondrial dynamics in mammalian cells. Biophysics Reports, 3(4–6), 64–72. https://doi.org/10.1007/s41048-...

...sub-cellular localization signals, plant markers, and reporters (Engler et al., 2014). This kit is a great starter...into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing of sgRNAs...generic MoClo system for any given species can be repurposed for CRISPR applications (gene insert = Cas9 +...Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media...

...Live-cell labeling strategies. The green structure represents a fluorophore or other useful ligand, gray structure... self-labeling tools: Tetracysteine: The first reported chemical labeling tag, from Roger Tsien’s lab ...self-labeling fusion tag based on the human DNA repair protein AGT that gets covalently linked to derivatives...collection. CLIP-tag: Also in 2008, Kai Johnsson’s lab reported another AGT-based tag named CLIP-tag, which reacts...

...Clustered Regularly Interspaced Short Palindromic Repeats: Sequences of DNA found in bacteria that allow ...CRISPR can be used to cut the DNA sequence. With a repair template, CRISPR can be used to introduce a functional...deaminase can convert adenosine to inosine. Activate/Repress: Modified versions of CRISPR nucleases that can...

...grant proposals or to demonstrate impact in annual reports. While the raw data is, of course, interesting ...data, including total requests, total countries represented by requestors, year of first reagent sent, year...and the source code is available in the Github repository (you can find the link in the upper right hand...