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Showing: 561 - 580 of 1071 results
  1. GCE4All: Making Genetic Code Expansion Accessible

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    ...trade-offs. This balance can be extremely tricky to find: each of the factors can negatively impact both...or chemically reactive side chains, all of which can help determine structure and function relationships...relationship between structure and function…if you can get your protocol to work. Or, to quote Ryan Mehl...robust and universal techniques that researchers can easily implement in their own labs. They provide ...bulletin board on their website specifically so people can talk about GCE. In fact, their first protocol is ...proteins, you’re probably quite interested in the canonical 20 amino acids. But in your quest to learn more...code expansion (GCE): techniques in which non-canonical amino acids (ncAAs, also sometimes called unnatural...
  2. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

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    ...publication describing how short term storage on dry ice can shift sample pH, we wondered whether or not the dry...affected by dry ice, and specifically how dry ice can affect virus from Addgene. Bottom line: there’s ...temperatures (-80°C) and once in the tube, the CO2 can alter the pH of the liquid sample. The good news ... that this effect is reversible and the pH shift can be prevented. Keep this information in mind if you...C. For samples that are shipped on dry ice, this can affect the pH of the sample, which may have some ...Additional Resources on the Addgene Blog Learn How You Can Use the Viral Service Tips for 1st Time CRIPSR User...
  3. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

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    ... indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured. Archon1...rapidly screen thousands of proteins for one that can report electrical activity via imaging. They took...nature, but developing them into optogenetic tools can be difficult because this requires detailed structural...interactions in cells because the protein binders can be customized for protein visualization, relocation...these photo-reversible protein-protein interactions can control subcellular localization processes in vivo...
  4. How to Keep a Lab Notebook for Bioinformatic Analyses

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    ...that lays out which experiments can be found in each folder. This can be a simple .txt document. Record...addition to output files, this is also where you can store any power point, excel, or other files relevant...the camelCaseFormat since spaces between each word can make it difficult to accurately call a file. Version... track of your exact changes. Alternatively, you can simply make a note in your methods section about ... Lab Notebook Organized Learn about Software You Can Use to Save Time and Track Your Experiments Try out...
  5. New Tool for Lineage Tracing: The ClonTracer Library

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    ...clone can then be monitored over time by sequencing the barcodes in the population (all barcodes can be ...of cells with specifc, easily read sequences that can be used to measure their clonal abundance before ...tumor xenograft models. She hopes that the library can be used to identify appropriate, non-overlapping ...development - such that initial resistant populations can be eradicated up-front quickly. This approach is ...25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct evolutionary paths to become resistant...resistant cancer cells exist prior to drug treatment. They infected a non-small cell lung cancer cell line...tool to look at a variety of questions surrounding cancer development, metastasis, and drug resistance, Bhang...
  6. CRISPR 101: Non-Homologous End Joining

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    ...smegmatis (Yan et al., 2020). NHEJ can also be engaged by variants of the canonical Cas9 approach. A pair of CRISPR...system for genome engineering is the fact that Cas9 can be easily programmed to make a DNA double strand ...frameshift mutation. Of some importance, the deletion can be less heterogeneous when constrained by sequence...loading of Ku and normal NHEJ activity. Other factors can also influence NHEJ activity. For example, repressing... flank regions of hundreds of base pairs or more can simultaneously introduce a pair of chromosome breaks...containing compatible overhangs is available. Cas9 can also be altered to generate a targeted single strand...and Biophysics and the Lineberger Comprehensive Cancer Center at UNC Chapel Hill.   References Bennardo...
  7. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

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    ...victoria the functionalized nanobodies described here can be used to target any protein fused with GFP, YFP...vicinity with biotin. The biotin-labeled proteins can then be isolated and analysed. In this work, the ...School have developed a group of plasmids (here) that can be used to label all proteins within a radius of ...Tyrosine) with biotin. These biotin-labeled proteins can them be analyzed by mass spectrometry allowing a ...composition of proteins found at the telomere, and hope it can be used to expand our understanding of proteome localization...MarathonRT, isolated from Eubacterium rectale, significantly outperforms other RTs in this regard and is ...
  8. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

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    ...Additionally, you can upload your own images to the Biorender workspace. Free Biorender accounts can store 5 workspaces...color and size of icons are adjustable. New icons can be requested, although there’s no guarantee Biorender...informal settings like lab meeting presentations. You can download larger images without the watermark, which...then clicking the “Usage rights” dropdown menu, you can search for images “Labeled for reuse with modification...Google Image Search results includes images that can’t be reused without attribution, despite filtering...
  9. New and Upcoming Viral Vectors - Spring 2019

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    ... of the AAV we have released so far in 2019. You can also browse our entire AAV inventory! Our new AAVs...fluorescence of the GCaMP7 family of calcium sensors and can be used for imaging of small neuronal processes like...Since it’s illuminated by red/orange light, Archon1 can also be used in conjunction with optogenetic activators...AAV These fluorescent protein expression vectors can be used to visualize cells and to quantify targeting...expression to GABAergic interneurons. These vectors can also be used as the control vectors for experiments...
  10. Sleeping Beauty Awakens for Genome Engineering

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    ...sequences of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both ...occurs which cuts the DNA.3.) The released transposon can now bind a strand of DNA with a TA dinucleotide (...transposon containing the gene-expression cassette that can translocate within the genome. The Sleeping Beauty...generation of stable transgenic cell lines which can robustly express your gene-of-interest, either constitutively...Additionally, in 2014 researchers at MD Anderson Cancer Center presented results from a clinical trial ...
  11. Hot Plasmids: Summer 2024

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    ...editing and twin prime editing can integrate DNA cargoes of 80–100 bp, PASSIGE can integrate cargoes over a ...concentration and purity of the target molecule, which can be problematic for low-abundance targets or natively...plunge freezing By Rachel Leeson Cryo-EM samples can be damaged at the air-water interface (AWI) during...small proteins that naturally adsorb to the AWI, can protect cryo-EM samples from damage (Abe & Lim, 2024...air-water interface (AWI) before vitrification, they can be completely denaturated (left) or complexes may...of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing (Nuñez et...methyltransferase for epigenetic silencing, which can be toxic, CHARM uses a cofactor fused to a histone...
  12. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    ...CRISPR components for plant genome engineering CRISPR can be used to knockout, activate or repress target genes...required. As with other model systems, multiple gRNAs can be expressed to modify several genomic loci at once...dCas9-VP64) or repressors (dCas9-KRAB or dCas9-SRDX) can also be used to activate or repress target genes ...result in a failed experiment. CRISPR components can be expressed stably or transiently depending on the...method and cell type in question. CRISPR components can be delivered and expressed transiently using a standard...information on Agrobacterium-mediated transformation can be found in this blog post. The pDGE Dicot Genome...transformation in plant cells. Agrobacterium tumefaciens can be used as a vehicle to deliver your gene of interest...
  13. CRISPR/Cas9 FAQs Answered!

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    ...How can I quickly check if the cloning of my oligo into a backbone like PX330 worked? A12: You can quickly...newest CRISPR technologies and their applications can be exhausting. A quick search for "CRISPR", short-hand...concept of the double nickase system is that you can express two different chimeric gRNAs with the Cas9... cuts 3–4 bp upstream of the PAM sequence. There can be some off-target DSBs using wildtype Cas9. The ...Tips for using homologous recombination (HR) Q7: Can you summarize HR template vector design guidelines...shorter. Q10: What is the maximum amount of DNA that can be inserted into the genome using CRISPR/Cas for ...reference for using ssODN as HR Template? A11: You can take a look at Soldner et al. (2011) which uses ssODN...
  14. Designer PUF Proteins for Any RNA Target

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    ...binding factors (PUF). In a single step, researchers can now assemble designer PUF domains for RNA specificity...interesting because by changing certain amino acids you can change the specificity,” explained Zhanar Abil of...repressor. In principle, however, designer PUF proteins can be fused to other components to lend them many other...change the intracellular localization of RNA. “You can think of changing bases to other RNA bases or unnatural..., for example, by targeting pathways underlying cancer or other disease processes. Get the PUF Assembly...
  15. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

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    ...plants. In the most practical terms, what GreenGate can do is “save you half a year,” in comparison to more...fragments produced will have different overhangs, they can be designed such that they go back together only ... restriction endonuclease of choice, BsaI, which can be very sensitive to temperature fluctuations. He...sensitive protein,” he says. “There is nothing you can do about it but be gentle. If it no longer works,...
  16. Let There Be LITE Plasmids

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    ...within minutes of light exposure. Third, the response can be graded with light dose and is fully reversible...reversible after light retraction. Finally, because light can be applied non-invasively, its use is not restricted... endogenous cellular processes." The LITE system can now be applied to unraveling the genetic and epigenetic...time by making an AAV1 supernatant. (The details can be found here.) So tell us, readers, how will you... the brains of living mice both quickly and significantly. Genes under LITE control showed a rise in transcription...
  17. Overwhelmed? Take a Break With Our 5 Favorite Science Comics

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    ...extra push, there’s evidence that laughter really can be good for you. A search on PubMed for the word ... few fun and very recent bits of evidence. (What can I say, we are scientists). Researchers in Taiwan ...those young people in a better mood. Laughter itself can sometimes even count as demanding exercise, and, ...Taiwan showed that a laughter program significantly decreased immunological markers of stress in adolescents...
  18. 22 Hot Plasmid Technologies from 2014

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    ...2) which can be utilized in three successive assembly steps. Before beginning, scientists can insert fragments...lab has engineered a modular mini-Tn5 vector that can be used to generate random mutagenesis libraries ...bacterium Pseudomonas putida and demonstrate that they can successfully create GFP fusion proteins with a variety... cytochrome c oxidase subunit VIII, so that they can be imported to the mitochondrial matrix. Gottlieb...pathway or induce other compensatory mechanisms. How can protein function be dissected while minimizing confounding... a membrane-permeable pharmacological agent that can rapidly induction heterodimerization of proteins ...the final construct. Up to eight entry vectors can be used for each Multisite Gateway compatible destination...
  19. Viral Vectors 101: The Retroviral Lifecycle

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    ...sort of virulence as well — namely that they can cause cancer via insertional mutagenesis (which HIV does...simple spokesmodel for the whole family; however, you can use Figure 1 to compare the genomes of these two ...particle — and of the normal, replicative life cycle — can help you understand these seemingly odd choices and... of its first steps! However, retroviral vectors can still direct the translation of proteins because ...they have two dimerized linear ssRNA strands (which can have a different genomic sequence from one another...et al., 1971) and the full wild-type viral genome can direct the production, packaging, and release of ...viral particle, and how we as dedicated researchers can exploit it. This blog was written by Addgenie Brian...
  20. Fluorescent Proteins 101: Fluorescent Protein Timers

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    ...monitoring localization; using FP timers, researchers can look at a single image of a cell and understand how...correlating this ratio to the total expression time can therefore be used to determine how long a pool of...for the theoretical timer shown in Figure 1B, you can use the equation fit to this linear region to solve... one benefit of the common FP timers is that you can monitor age by expressing a single FP-timer protein... oxygen, temperature, and pH, but timer activity can be calibrated to these different conditions, enabling...aggregation-prone tetrameric fluorescent proteins can be. Cytotoxicity, improper localization, and decreased...circuit (Saxena et al.) In a general sense, FP timers can be used in any situation where one wants to understand...
Showing: 561 - 580 of 1071 results