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We narrowed to 993 results for: Des

Showing: 241 - 260 of 993 results

  1. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...optimized. How should I design my mutagenic primers? The key is to design primers that are the same ...generation of a plasmid library that expresses all desired variants of a protein. Applying a selective pressure...PCR that uses mutagenic primers to introduce all desired codon changes (step 1 in the image above). A second...ssDNA is packaged into phage and then extracted. Besides taking a couple days to generate the phage-derived... The one-pot saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach for ...Treating the plasmid with ExoIII and ExoI then fully degrades the nicked strand. Synthesize mutant strand: A...

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

    Type
    Blog Post
    ...any DNA sequence in the way that they like, the design and engineering of fusion proteins is relatively...fusion proteins and highlight some aspects of their design.  Protein size and shape matters The green fluorescent...necessary for post-translational modifications. Despite these potential issues, GFP has been used successfully...consist of random amino acid sequences or (ii) designed to form an inert, unstructured peptide and therefore...C-terminus of the fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus...2011).  Although structural information may guide design, it is advisable to generate multiple constructs...over-expression artifacts The introduction of DNA that encodes a fusion protein adds protein to the existing pool...

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

    Type
    Blog Post
    ... While it is certainly possible to select for desirable traits using traditional plant breeding approaches...modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR has already been used...wheat, mushroom, and rice (Khatodia et al., 2016). Despite the almost universal functionality of the CRISPR...genes in plants using the same general experimental design principles developed in other model organisms (...nanostructures (Zhang et al., 2019) or cell-penetrating peptides (Santana et al., 2020) and plants viruses (barley... through Addgene. In addition to the plasmids described above, Addgene carries several useful CRISPR toolkits...CRISPR Multiplexing Tools Validate Your Genome Edit Design Your gRNA Additional Resources on Addgene.org ...

  4. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...comments section.  FX cloning kit This kit is designed to be used with Type IIS restriction enzymes and...this kit have been used to purify a variety of peptides including antibiotic efflux pumps and ankyrin ...et al. "A transcription blocker isolated from a designed repeat protein combinatorial library by in vivo...followed by leader sequences (encoding small leader peptides) downstream of which you can clone your gene of...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to...(2015): 31-37. pCri system The Gomis-Ruth lab designed the pCri system to simplify the protein purification...for cloning. Furthermore, some plasmids contain peptides to facilitate protein folding and/or protease ...

  5. CRISPR Plasmids - Parasites

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    Collection
    ...genome-wide knockout gRNA pooled library designed with 10 guides against 8,158 predicted protein-coding ...CRISPR expression plasmids that have been designed for use in parasites. CRISPR...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Eukaryotic parasites continue...have been the focus of biomedical research for decades. Recent developments with CRISPR-Cas9 gene editing...

  6. Institute for Protein Innovation

    Type
    Collection
    ...signaling during development. The IPI collection includes anti-glypicans that accurately detect glypican... Glypican Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Integrin... Integrin Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Browse...common cellular proteins. ID Recombinant Antibody Description Isotype Reactivity Recommended Applications Browse...express plasmid-based antibodies. ID Plasmid Description...

  7. CRISPR Plasmids - gRNAs

    Type
    Collection
    ...Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA plasmids designed to target various...pyogenes , S. aureus etc.) was this gRNA sequence designed for? A given gRNA sequence may only be compatible...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, PAM requirements, experimental...unpublished plasmids containing gRNA sequences designed to be used with any CRISPR nuclease and function...

  8. A Control for All Seasons

    Type
    Blog Post
    ...controls and be able to use this information to design thoughtfully and thoroughly controlled experiments...is generally a sample or group that will have a desired response. For example, if you are studying the ...sample, the antibody binds specifically to its desired target (purple) and nonspecifically through the...The Preimmune serum control will not bind to the desired target but will bind nonspecifically through the...the Fc, allowing you to detect this undesirable interaction. When non-specific interactions are suspected...nonspecifically with positively charged groups, such as aldehydes in fixed samples. To address this, block positively... will be a useful starting point for you when designing rigorous experiments. While this blog touched ...

  9. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    ...most clinically-relevant proteins are also the hardest to study. One common strategy is protein overexpression...compatible with both strong and weak promoters, and even modest endogenous pools generate high signal-to-background...introduce mutations. This vector-based use of HiBiT provides a means to quickly answer research questions before...the code of 'undruggable' targets Researchers can design drugs to bind to well-defined ligand-binding pockets...mechanistic drug responses (Kamio et al., 2025). They designed HiBiT-tagged TP53 constructs, one with an R175H... links p53 mutations to treatment outcomes and provides a HiBiT-enabled framework for quantifying variant...many difficult-to-study protein targets, HiBiT provides an excellent solution compatible with high throughput...

  10. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren Looger and Kaspar Podgorski...Aggarwal et al., 2025).In this latest work, the team designed and screened a large library of iGluSnFR3 mutants... HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's what makes these... stable cell line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked... plasmids for flexibility and speed, so you can design, test, and innovate faster than ever. Plus, we'...have a two-for-one solution: TurboCas!TurboCas provides the genomic targeting power of dCas9 with the ....2024.11.007. Branon, T. C., Bosch, J. A., Sanchez, A. D., Udeshi, N. D., Svinkina, T., Carr, S. A., Feldman, J. ...

  11. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional control...Organoid EasyTag workflow combines thoughtfully designed plasmid components with Fluorescence Activated... heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating... rather than on the fluorescence intensity, it provides a direct and quantitative detection of SLs. The...biosensors could have a major economic impact.   Design and mechanism of action of the receptor-based SL...targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize on-target cleavage and...

  12. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ... Roth BL (2015) DREADDs (Designer Receptors Exclusively Activated by Designer Drugs): Chemogenetic Tools...less invasive and more flexible as it relies on designer ligands that can be simply introduced by injection...lab and Takafumi Minamimoto’s lab known as deschloroclozapine (DCZ) has been developed to meet these requirements...should be carefully controlled for in experimental design.   Optogenetics Chemogenetics Timing Fast ...Find our chemogenetics and optogenetics science guides ...

  13. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...tagging method that labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6...Direct reprogramming describes the process where differentiated cells are turned into a cell type of ...with the majority of cells failing to develop the desired identity. The development of single cell technologies...Morris Lab at Washington University in St. Louis provides a combinatorial indexing strategy that simultaneously...towards successful iEP reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. ...

  14. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ... been described in this blog. If your data does not meet the assumptions for the tests described above...experimental groups, but how do you determine if they deserve that oh-so coveted asterisk of significance? You...referred to as two-sample or independent, but these designations all refer to the same test. On some occasions...Correction and the Benjamini-Hochberg procedure are designed to reduce false positive rates and control for...error rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical... making your statistical analysis plan when you design your experiment, instead of after you’ve already...

  15. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    ...or FP) is one of the most important steps in the design of an imaging experiment.  The choice is non-trivial...fusion protein, etc...).  There are many online guides – including an excellent series of posts by Joachim...information about fluorescent proteins.  It was designed with the goal of encouraging user contributions...while also maintaining a relatively strict database design (schema).  Registered users can submit new fluorescent...risk of inaccurate data submissions, so FPbase includes a versioning system that allows users to see when...for, the Table of Fluorescent Protein Properties provides a master list of all proteins in the database,...

  16. Science Career Options

    Type
    Blog Post
    ...to a wide range of industries and is a highly desirable quality sought after by employers. These skills...correctly the first time. These skills are very desirable to employers because they save time and money ...Meteorologist Journal Editor Textile Technologist/Designer     Fieldwork and traveling Many people are...employers. Increasingly, computer skills are highly desired, as roles are becoming more reliant on technology...(Python, R, SAS, HTML, Matlab etc) are highly desirable in a range of roles, including: Science Reporter...find answers to include: Will you be sitting at a desk all day or be on your feet constantly in a lab? ...

  17. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    ... lacks context unless it links to a verified description of that reagent online. Furthermore, if the link... link between the catalog number and reagent description is lost (e.g. a supplier is bought out by another...directly to the original article online. It is also designed to be persistent and exist online indefinitely...specific information about the plasmid: name, description, how the plasmid was generated, validated sequence...authors comply with journal policies, Addgene now provides an RRID and a resolvable compact identifier for...you directly to the lenti-Cas9-VQR plasmid page. Despite their differences, both of these PID formats for...

  18. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...lentiviral packaging methods. The three components described above (envelope, packaging, and transfer) are ...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...nomenclature of the plasmid backbone is meant to be descriptive of the components found within the backbone. ...

  19. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ... has precluded widespread adoption of the technology. In 2012, Saito et al. described the first Nano-...molecular and cell biologists over the last two decades, but their use can be limited by a few caveats....hours! The latest enhanced Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous...substrate furimazine. In addition to the previously described applications, the enhanced signal of eNL allows...autofluorescence and a high signal-to-noise ratio. Despite the many benefits of Nano-lantern technology, there...

  20. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a  screen comes in. Let’s...blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse...Sanger sequencing determines the precise order of nucleotides within a DNA molecule, in this case a plasmid... of the information you need, so you'll want to design a custom primer. Because Sanger sequencing can ...
Showing: 241 - 260 of 993 results