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We narrowed to 993 results for: Des

Showing: 101 - 120 of 993 results

  1. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ....  These primers are typically ~70 nucleotides long (20 nucleotides that anneal to the DNA sequence of..., the substrate should be ~70-100 nucleotides long with the desired alteration(s) located in the center...genomes. 2) Designing ssDNA oligos that avoid MMR activation:  There are two ways to design oligos that...tube. The donor DNA substrate only requires ~50 nucleotides of homology to the target site for recombination... be explained in greater detail. Substrate DNA design and generation Expression of lambda red recombination...antibiotic resistance cassette.   Substrate DNA Design and Generation Whether you use a linear dsDNA or...insertions or deletions greater than approximately 20 nucleotides, while ssDNA substrate is best for point mutations...

  2. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...preprint title or a general description of the deposit. Select your desired distribution status: Hold...choose from one of the following descriptions for your plasmid: Encodes insert(s) A plasmid with one or...should describe how your plasmid is intended to be used. For instance, if your plasmid is designed to allow...Unpublished or pre-publication plasmids" and set a desired distribution status. You can submit "Published ...plasmids with the original publication where you described them. Search for your article as you would search...distribute, or Addgene finds your journal publication describing the plasmids, or (for deposits submitted after...selected your publication or entered your deposit description, the next page will allow you to add plasmids...

  3. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...template vector design guidelines? A7: A few notes below are considerations for designing HR donor. Generally...usually pick 3–6 guides around the region to find the most efficient guide (most guides would work, but... this ‘split RNA’ design is not as efficient in genome editing as the second design (in pX330/PX335), ... CRISPR plasmid tools and numerous experimental design decisions to make, it makes sense that scientists...CRISPR forum's most popular questions and answers. Designing your CRISPR genome editing experiment Q1: Should...comparable efficiency to the optimized chimeric design but with better accuracy (in other words, lower...nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong et al. (2013) Science ...

  4. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...list of Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism...website, which breaks the design process down into four easy steps, provides the real value for the user...potential of them not working, or spend a few weeks designing and cloning the vectors you actually want. But... Fragmid is a modular vector assembly process designed for a quick, plug-and-play approach to cloning...et al., 2024). It reduces the amount of time to design and create a new vector to four days for an experienced...high school interns, were able to use Fragmid to design and clone their vectors in five days. Did you say...s modular assembly structure takes a lot of the design work out of the plasmid creation process. Each ...

  5. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ... pii: 56469. Edwin Chapman Neuropeptides GRAB biosensors for neuropeptides VIP, NPY, CCK, NTS, UCN, SST... orexin neuropeptides A genetically encoded sensor for in vivo imaging of orexin neuropeptides. Nat Methods...Biosensors Fluorescent biosensors are proteins designed so that their fluorescent properties are sensitive... or organisms for long-term imaging and can be designed to specifically localize to cellular compartments...compartments or organelles. Addgene’s database includes many fluorescent biosensors that signal the presence ...environmental conditions. With proper experimental design, biosensors can enable investigation of a signaling... on Request for many more. Metal Ions Target Description Article PI Calcium jGCaMP8 Fast Genetically Encoded...

  6. Plant Plasmids and Resources

    Type
    Collection
    ...contains many plasmid tools that were specifically designed for use in plants. These resources include tools... to fluorescent proteins. Addgene’s collection includes plasmids containing promoters that drive general... Chen. Cas9 and gRNA backbone for use in rice. Includes a protocol (Link opens in a new window) for Targeted... Yang. The MoClo CRISPR/Cas Toolkit for Plants includes CRISPR/Cas nucleases, base editors, gRNA backbones...Nekrasov. The MoClo-compatible zCas9i Cloning Kit includes a high efficiency intron-optimized SpCas9-coding...Caixia Gao. Cloning The following collections are designed for creating plant expression vectors: A modular...MoChlo: Modular Cloning Chloroplast Toolbox for design of synthetic operons for chloroplast transformation...

  7. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...ligated into Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt...cell that gets the plasmid contains all of the desired gRNAs. This increases the chance that all the edits...promoters. To use this plasmid, you simply order oligonucleotides for your chosen gRNA target sequences and ...ordered assembly of multiple components into a destination vector. The first step in CRISPR/Cas9 Golden ...Golden Gate multiplexing is to clone the oligonucleotides specifying each gRNA target sequence into distinct...together to allow for ordered assembly into a destination vector that expresses Cas9. This is illustrated...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...

  8. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...primer design Design primers for Goldengate cloning   Design primers for In-Fusion Cloning Design primers...traces, while the paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While...academic users. Serial Cloner: Free desktop-based software for plasmid design and mapping. ApE (A plasmid Editor...Primer design tools Many of the above-listed plasmid mapping tools have built-in primer design tools, ...stand-alone sites for general primer design, as well as primer design for different cloning applications...characteristics. NEB Data-optimized Assembly Design (DAD): This tool designs overhang sets for high complexity Golden...Primer design. Plasmid mapping. DNA sequence analysis. We all have our favorite tools for tackling these...

  9. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate reporter gene... can't give all of them attention they righlty deserve, but, in this post, we'll provide a small sampling...-2xNLS-VP64 and dCas9(N)-FRB-NES). This system provides users greater temporal control over CRISPR/Cas9...cell events Three years ago, Brian Kuhlman’s lab designed photoswitchable dimers by fusing the SsrA peptide...the authors recently used computational protein design, phage display and high throughput binding assays...transcription of downstream genes. Nihongaki et al describe the targeted activation of endogenous genes and... a gene-specific forward primer containing the desired restriction site (the MCS has several options) ...

  10. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...customized to direct assembly of DNA fragments. When designed correctly, the recognition sites do not appear... scheme is as follows: the gene of interest is designed with Type IIS sites (such as BsaI or BbsI), that...and do not appear in the final construct. The destination vector contains sites with complementary overhangs...ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a single...IIS enzyme and ligase. Although the original destination vector + insert may spontaneously religate, this... be re-digested. In contrast, formation of the desired ligation product is irreversible because this construct...a single reaction! These overhangs specify the desired order of fragments, and the loss of enzyme recognition...

  11. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ... reference?  The CRISPR guides used in the screens they analyzed were designed to have a perfect match...African descent have approximately 20% more single nucleotide polymorphisms (SNPs) that map to guides in the...accounted for in the CRISPR guides they used, leading to mismatches between the guides and the lines’ genomes...modify the CRISpick algorithm to prevent guides from being designed that target regions with high genomic...coming through an unexpected source: the CRISPR guides, short pieces of RNA that bind to a cell’s DNA,...x-axis) which SNPs map to thetargeting sequences of guides across 8 CRISPR libraries. Samples are divided ...are more common in individuals of African descent.Correcting the bias  While their thorough analysis uncovered...

  12. Worm Expression Resources

    Type
    Collection
    ...the biology of C. elegans and other nematodes. Wormbuilder - Describes methods to engineer the genome of...characterized which allow researchers to express desired constructs in precise cell types and/or developmental...that contain tools for use in worm research. This includes CRISPR, RNAi, empty backbones, fluorescent proteins...expression plasmids. Large collections or kits are described below and not included in this table. For a more...search Addgene's entire collection. ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes...strains. CRISPR plasmids from John Calarco's lab as described in Heritable genome editing in C. elegans via ...genome-editing events. Developed in Craig Mello's lab and described in A Co-CRISPR Strategy for Efficient Genome Editing...

  13. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ...suspense. Notice the simplicity of their slides and how the slides complement the spoken message. Learn from...speak to start creating slides or thinking about content. Create a Google.slides or Powerpoint file right...how these speakers talk without looking at their slides or notes. Listen to their stories and narratives...listeners to be thinking “What happens next?”  Describe how you pushed to a solution or overcame obstacles...term, but are worried that this might sound condescending, consider explaining the term on a slide but...talk will really take and to adjust content and slides to better suit the target time frame.   I suggest...peers than a less supportive audience? 7. Hone Your slides Use big images that are easy to see (and for heavens...

  14. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...experiments that can be partially designed using the Snapgene desktop software. If a plasmid contains a...sequencing results with reference sequences To design experiments for further plasmid verification Identifying... contents of the resulting FASTA file into the desktop version of Snapgene to visualize the sequence and...in Snapgene’s extensive database. Snapgene also provides easy visualization of open reading frames, which...any known sequences for the insert. The Snapgene desktop software allows us to quickly align multiple sequences...verification experiments: Primer and restriction digest design Some regions of plasmids are difficult to sequence...region with a high amount of guanine and cytosine nucleotides, we might need to send it for Sanger sequencing...

  15. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...experimental designs and we need to be transparent about the reasons we alter these designs if we do so...experimental design to a journal prior to performing the experiment (or set of experiments), the design would...from the event can be found below along with a description of the program beneath it. You can jump to the...the different time stamps in the description to watch any sections you’re particularly interested in, ...JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier...and not just impact. Cause #3: Poor experimental design It’s easy to get excited about positive results...experiments to work, but part of good experimental design is recognizing that we’re biased and taking steps...

  16. CRISPR Plasmids and Resources

    Type
    Collection
    ...Bacteria CRISPR plasmids designed for use in bacteria. C. elegans CRISPR plasmids designed for use in C. elegans... CRISPR plasmids designed for use in Drosophila . Mammalian CRISPR plasmids designed for use in mammalian...Parasites CRISPR plasmids designed for use in parasites. Plant CRISPR plasmids designed for use in plants. Yeast...Yeast CRISPR plasmids designed for use in yeast. Xenopus CRISPR plasmids designed for use in Xenopus . ...Browse depositor protocols, find software for gRNA design and deep sequencing analysis, discover links to.... Zebrafish CRISPR plasmids designed for use in zebrafish....

  17. Lentivirus Plasmids

    Type
    Collection
    ...plasmid. Cheng 17452 pLenti CMV Puro DEST 3rd Gateway destination plasmid with constitutive or inducible...lentiviral systems available at Addgene have been designed for increased researcher safety. Read our Lentiviral... are used in conjunction with the plasmid that encodes the viral genome (typically called the transfer...lentivirus technologies. ID Plasmid Generation Type Description PI 12253 pRSV-Rev 3rd Packaging Contains Rev;...for research, it is the lentiviral genome that encodes genetic material that the researcher wants delivered...table and search "lenti". ID Plasmid Generation Description PI 8453 pLKO.1 puro 3rd U6-driven shRNA empty...cloning plasmid 3rd U6-driven shRNA empty plasmid; includes a stuffer for easy cloning Root 14748 pLKO.3G ...

  18. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ... (V70) (Plasmid #188537) . These reagents are described in detail in Mohler et al. (2023) and help deliver...purifying the phosphosite library as GST-fusion peptides (“Mode #1” expression). The Mode #1 phosphosite...Phosphorylated Isomers (iSPI) The iSPI resource described in Galloway et al. (2022) is adapted from the ...the synthetic human phosphoproteome. It subdivides the 110,139 human phosphoserine site library (Pooled ...Iterative Synthetically Phosphorylated Isomers (iSPI) described in: Gassaway, B. M., Li, J., Rad, R., Mintseris...in a new window) rEcoli XpS and a new pSerOTS described in: Mohler, K., Moen, J. M., Rogulina, S., & Rinehart..., 551 KB) Human Serine Phosphopeptide Library described in: Barber, K. W., Muir, P., Szeligowski, R. V...

  19. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ... massive amount of descriptive transcriptomics data. Custom CRISPR screens designed to target differentially...Institutet, in Sweden. It can be very time consuming to design 5 guide RNAs (gRNAs) targeting each of the 1000...). Green Listed is a new software tool used to design gRNAs for custom CRISPR screens targeting a (long...moment, 13 full genome SpCas9 knockout libraries designed by different academic labs, targeting mouse, human...drug targets  Another interesting approach is to design screens against known druggable targets, preferentially...library from Addgene, and then use Green Listed to design a smaller validation screen using selected gRNAs... could also be combined. For example, you could design a screen targeting all differentially expressed...

  20. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...often use the Cre/Lox system described earlier. In this case, you design your targeting vector such that...replacing it or disrupting it with an piece of DNA designed in the lab. Specially constructed plasmids can...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate...inactivate specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems...locus with homologous sequence using a specially designed vector that contains sequence homology to the ... the process, we’ll walk through an experiment designed to knock out exon 2 of a given gene.   ... Figure 1: A knockout targeting vector designed to insert a resistance gene. The vector contains a neomycin...
Showing: 101 - 120 of 993 results