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We narrowed to 634 results for: des.2

Showing: 101 - 120 of 634 results

  1. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ...CRISPR to detect SARS-CoV-2, including many with plasmids deposited at Addgene! 2. DNA Purification Without...detection methods, synthetic nanobodies, SARS-CoV-2 protein expression, and more. We also highlighted ...empty backbones, fluorescent protein tags, SARS-CoV-2 plasmids, etc. It is our hope that by providing cloning...RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas by ...databases, DNA sequence manipulation, and primer design. Best wishes for the new year and your research...

  2. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    ...Rodrigo Arturo Rivera Solís, Universidad Autónoma de Yucatán, Mexico Dr. Rodrigo Arturo Rivera Solí... genetically encoded H2O2 probe is pQE-30-HyPer3 (2) which he will co-express together with a plasmid ...Pharmacology 2017 Jun 15;134:1-17. PubMed PMID: 27984002. 2. Bilan, Dmitry S. et al. “HyPer-3: A Genetically Encoded...renewable biotechnology products and biosensors for herbicides as well as developing yeast as source for dietary...technology to engineer plants that are able to destroy DNA viruses, similar to the anti-phage CRISPR effect...cloning kit) she intends to make yeast capable of desaturating fatty acids. Identification of Insecticidal ...as insecticidal proteins, the use of chemical pesticides may be avoided or decreased. Biosensors for herbicide...

  3. Hot Plasmids - October 2022

    Type
    Blog Post
    ...and 2), they found an indicator that is optimized for 2-photon imaging of voltages. This includes monitoring...resultant JEDI-2P sensors overcome many challenges in 2-photon in-vivo imaging of voltage fluctuations. Image...suitable for in vivo optogenetic use.   Fig. 2: A) HcKCR1 is highly selective for K+ over other Na...slice of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential frequencies...KCRs), from a fungus-like protist Hyphochytrium catenoides (Govorunova et al. 2022). The authors show that...

  4. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...antibody has an antigen binding site for a total of 2 binding sites per antibody. C) IgG’s are big enough...enough that multiple labels (green circles in figure 2 below) can be attached to each antibody. But being...displacement Require separate incubation of 1° and 2° Must use different species and/or different IgG ...Greater resolution due to lower label displacement 2° incubation can be skipped by pre-binding to 1° ...a fluorescent tag and the target antigen to about 2 nm, which can lead to higher resolution for super-...subclass.   Express the Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express... Find plasmids from this publication at Addgene. 2. Bates, M., Böhning, M.I., Chug, H., Görlich, D., ...

  5. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ...sequence YFP Thorben Dammeyer 54520 mCherry-Peroxisomes-2 Peroxisome Peroximal Targeting Signal 1 (PTS1) mCherry...Centrin2 Centrioles (dependent on cell cycle) Centrin-2 EGFP Erich Nigg 41151 pEGFP Cep170 C-term Centrioles...Robert Singer 27382 pDEST/N1-hEB1-GFP Microtubules EB1 GFP Robin Shaw 40908 pDEST/LIfeAct-mCherry-N1 Actin...Davidson 40907 pDEST/hCx43-EGFP-N1 Gap Junctions Connexin 43 EGFP Robin Shaw 49861 pDEST-mCherry-GJA1-20k-N1...important to determine where your protein of interest resides. There are a variety of methods for determining...junctions, gap junctions, tight junctions, and desmosomes. Browse or search the table below to find markers...

  6. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...). A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons. Neuron, 92(2), 372...α2,3-/α2,6 N-linked sialic acids1 AAV receptor (AAVR)2   AAV2 Heparan sulfate proteoglycan (HSPG)3 ...3 AAV receptor (AAVR; not strictly required)2 Human fibroblast growth factor receptor 1 (FGFR1)3 αVβ5...AAV5 2,3-N-linked sialic acid4 AAV receptor (AAVR)2 Platelet-derived growth factor receptor (PDGFR...Heparan sulfate proteoglycan (HSPG)3 AAV receptor (AAVR)2 Epidermal growth factor receptor (EGFR)3 AAV8... AAV8 Laminin receptor (LR)3 AAV receptor (AAVR)2 Carboxypeptidase D (CPD), also called AAVR25   ... Terminal N-linked galactose3 AAV receptor (AAVR)2 Laminin receptor (LR)3 Putative integrin3 ...

  7. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...short-read sequencing are highlighted in Figure 2. Figure 2: Overview of short-read sequencing technology...software such as SPAdes, Shovill, apc, and NOVOPlasty. The pipeline contains three de novo assemblers,...manually checks and assembles the raw data.  Addgene’s de novo assembly pipeline usually generates the correct...raw FASTQ reads are used for either guided or new de novo assembly. If a full anticipated sequence is ... doesn’t resolve the assembly issues, we can redo de novo assembly within Geneious.  While this may seem...steps that gave us our initial incorrect assembly, de novo assembly within Geneious gives us additional...

  8. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly analyzed AAV-68544, we were...look for potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual.... When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing our...transduced cells whenever possible. This post will describe our workflow for viral genome sequencing (VGS)...

  9. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    .... Okita et al describes a protocol for generating iPSCs by plasmid transfection where 2 of 11 clones tested... cell reprogramming factors and wait for cells to de-differentiate. However it’s difficult, especially...plasmid-based expression requires serial transfection of 1 or 2 plasmids that express the reprogramming factors of...understanding the mechanisms of reprogramming, and 2) studies with clinical endpoints. In the first scenario...PubMed PMID: 27764162. PubMed Central PMCID: 5072705. 2. Sommer, Cesar A., and Gustavo Mostoslavsky. "The ...challenges." Journal of Cellular Physiology 228, no. 2 (2012): 267-75. PubMed PMID: 22767332.  3. Kim, Dohoon...PubMed PMID: 23546745. PubMed Central PMCID: 4176696. 2. Yusa, Kosuke, Roland Rad, Junji Takeda, and Allan...

  10. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ... that results in a polymerase error rate of up to 2% per nucleotide position. The “sloppy” PCR reaction...modifications: 1) increased concentration of Taq polymerase, 2) increased PCR extension time, 3) increased concentration...to synthesize a complementary mutant strand (step 2 and 3). Then the uracil-labeled template strand is...restrict site matter? Most of the time no, but there are 2 instances where it does. 1) When the backbone vector...ExoIII. See the below figure for an example like this. 2) When your gene of interest has a BbvCI site(s). If...saturation mutagenesis method are 1) ease of use, 2) time savings, and 3) ability to perform single site...PMID: 23766324. PubMed Central PMCID: PMC4409680. 2. Mccullum, E. O., Williams, B. A., Zhang, J., & Chaput...

  11. TALEN Plasmids and Kits

    Type
    Collection
    ...vectors designed for use with the Voytas Golden Gate TALEN Kit and are used in place of pTAL1, 2, 3, or...Sangamo)-Destination Pawel Pelczar pCAG-T7-TALEN(Sangamo)-Destination constructs were designed for optimal...pC-GoldyTALEN and RCIsricpt-GoldyTALEN are designed as destination vectors for the Voytas lab Golden Gate ...Plasmids pTAL5-BB and pTAL6-BB are designed as alternative destination vectors for the Golden Gate TALEN...TALEN(Sangamo)-FokI-KKR-Destination 40132 pCAG-T7-TALEN(Sangamo)-FokI-ELD-Destination Golden Gate TALEN 2.0...Mendenhall The TALE plasmid vectors described on this page are designed to target and edit the epigenome ...with custom repeat arrays. TALEN Kits TALEN Kit Description Depositing PI FusX TALEN Assembly System Contains...

  13. Plasmid Preps: Different Purity, Different Quantities, Different Uses

    Type
    Blog Post
    ... be extracted from small bacterial cultures (i.e. 2-3 ml) with or without commercially available kits ...ratios measured at 260 and 280 nm should fall ~1.8 - 2 and give scientists an indication of purity versus...contaminants. The 260 and 230 nm ratios should fall ~2 - 2.2, and provide an indication of purity versus ...hydrochloride used during plasmid extraction. Figure 2: Use cloning grade DNA for direct cloning of your ...other uses.   Cloning grade DNA Molecular cloning includes a set of techniques that are used to insert recombinant...

  14. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...non-homologous end joining (NHEJ) (2-5), depending on your desired genome edit. Since exploitation of ...each sample can be quickly calculated (details in [2]) and can be used to quantify rates of mutation in... have been reported for mismatch nuclease assays (2, 6, 7). Ability to distinguish between mono-allelic...to testing their performance (2). Remember that although digital PCR is a great screening tool, it still...PMID: 24509632. PubMed Central PMCID: PMC4063274. 2. Findlay, S. D., Vincent, K. M., Berman, J. R. & Postovit...your genome edits, this paper provides detailed instructions on how to design such assays, in addition to...great deal of precision. This means digital PCR provides an absolute measure of target abundance, does ...

  15. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...systems: a burst of class 2 and derived variants. Nature Reviews Microbiology, 18(2), 67–83. https://doi.org...online, searchable resource that provides summary information on Class 2 Cas enzymes. The information is...this content and much, much more.     Fig. 2 – List of resources and properties for a Cas entry...Database: A Functional Classification System for Class 2 CRISPR-Cas Enzymes [Preprint]. Biochemistry, Biophysics... is also documented. Experimental design When you are in the design phase of your experimental planning...is organized in wiki format and includes biochemical properties, previously established uses, and intended...relevant resources The homepage displays a graphic describing CasIDs and relevant properties of Cas enzymes...

  16. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ...them in Regenerative Medicine. Acta Naturae, 2010. 2(2): p. 18-28. PubMed PMID: 22649638. PubMed Central...capability to differentiate into specialized cell types [2]. We general speak of 4 levels of stem cell potency...zygote through to the morula stage are totipotent [2-4]. Pluripotency: When a cell has the ability to differentiate...stem cells (ESCs), which I discuss in detail below [2]. Multipotency: When a cell can differentiate into...PMID: 26726264. PubMed Central PMCID: PMC4686367. 2. Jaenisch, R. and R. Young, Stem cells, the molecular...essence, pluripotent cells can be thought of as descendants of totipotent cells. An example of pluripotent... produced Ethical concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation...

  17. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...Table 2 for more details on the effect of pH on the fluorescent properties of Rosella.   Table 2: Effect...in yeast. Autophagy,4(2), 205-213. doi:10.4161/auto.5331. PubMed PMID: 18094608 2. Sargsyan, A., Cai, J...environment like the vacuole or lysosome. See Table 2 for a summary of what color fluorescence Rosella emits...fluorescence accumulates in the vacuole. See Figure 2 for an example of what fluorescence looks like for... phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally PHO8 ...

  18. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...for the 2+ million DHS sites. See below for further details. DNA is a 3D Fractal Figure 2: DNA compaction... Find Enhancers that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG...expression for NANOG enhancers, we’re ready for step 2: design a CRISPR screen to disrupt these enhancers. The...NANOG enhancers 3. CRISPR out the enhancers from step 2. See which ESCs die or differentiate Step 3 reveals... CRISPR out the enhancers we identified in step 2. The result is a heterogenous population of stem cells...this article, Quilt CTO Aneesh Karve, shows how to design experiments that work anywhere in the genome. Aneesh's...enhancers! This leads us to a three-step approach for designing an experiment to identify the critical NANOG binding...

  19. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...different destination vectors depending on the total number of gRNAs you wish to clone, from 2-7. For example... plasmids allow expression of 2-5 gRNAs in zebrafish. Custom destination vectors are used depending upon...illustrated in the schematic below.   Figure 2: gRNA target sequences (colored rectangles) are cloned... plasmids: This plasmid set allows you to express 2-4 gRNAs, with four being the ideal number. First you...gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number). The 5’ most gRNA is ...Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through Golden Gate or Gibson Assembly. gRNAs...and a subsequent curing protocol that requires only 2-3 hours incubation. Kondo Lab multiplexed base editing...

  20. Fluorescent Proteins: FRET

    Type
    Collection
    ...referred to as intermolecular or bimolecular FRET) or (2) conformational changes within a protein where the...maturation time, and other environmental factors. When designing your experiment, be sure to refer to published...proteins fused with linkers of varying lengths. Description Article ECFP-EYFP FRET standards, fusion proteins...Biosensor Database (Link opens in a new window) designed by Eric Greenwald and the Jin Zhang Lab at UCSD...
Showing: 101 - 120 of 634 results