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We narrowed to 1,038 results for: LAS

Showing: 261 - 280 of 1038 results

  1. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...Penn State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid Prep Kit Amount 1000 Lanes...Two plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently...polyacrylamide gels. Get Tips on Verifying Your Plasmid Table 1: Comparing Typical Comericial Ladders...are significantly cheaper. When using a homemade plasmid prep protocol, it cost less than $10 to make 1000... 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30 per ladder...prep protocol here and the pPSU1 and the pPSU2 plasmids are available from Addgene. Learn How to Analyze...

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

    Type
    Blog Post
    ...background via P1 transduction. Plasmid Expressing lambda red genes from a plasmid allows for a mobile recombineering... flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA.  To use the lambda red recombineering...bacteria with integrated defective prophage 2) from a plasmid, 3) from mini-λ or 4) from the lambda red phage...phage pL promoter.  It is a good idea to choose plasmids that also express the repressors associated with...limit leaky expression of the Red system.  Using a plasmid to express the lambda recombineering system is ... do this is to express lambda red genes from a plasmid with a heat sensitive origin of replication.  Once... them at 42℃.  Mini-λ A hybrid between using a plasmid and stable integration of a defective prophage ...

  3. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ... labs and hinders experiments is mycoplasma contamination. Mycoplasma contamination has been shown to ...routinely test for mycoplasma contamination. There are several commercially available mycoplasma detection kits...larger microbes such as yeast, fungi, or bacteria, mycoplasma can be extremely hard to detect with levels reaching...cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients and can...the cell line’s growth and behavior. Signs of a mycoplasma contamination can be subtle and may include reduced...positive controls. Routinely test your cell line for mycoplasma and discard it if the culture has become overly...after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy is...

  4. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...loved helping to answer your plasmid and cloning questions through our Plasmids 101 series and by responding...Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids 101: Mammalian...Joanne Kamens Plasmids 101: Antibiotic Resistance Genes By Marcy Patrick Plasmids 101: Yeast Vectors...FAQs Answered! By Caroline LaManna Plasmids 101: What is a plasmid? By Margo R. Monroe Best wishes...as seen through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and...detailed information about this new technology. Our Plasmids 101 series continues to grab scientists’ attention...Genome Editing By John Doench (Guest Blogger) Plasmids 101: Origin of Replication By Kendall Morgan ...

  5. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...Chromatin-related genes with plasmids available from Addgene The links below pull up lists of plasmids containing the...containing lists of plasmids from the indicated publications. You can request all of these plasmids directly from...consolidate every conceivable chromatin-related plasmid in a collection that grows on a daily basis, the... does not have it. That said, if you know of a plasmid that we definitely don’t have that you really want...study chromatin and want to start sharing your plasmids through Addgene, don’t hesitate to start the process...these pages to learn when Addgene receives new plasmids containing these genes. As with the PIs above,... In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1 CREBBP CUL1 H2AFX...

  6. What's New in CRISPR - Winter 2018

    Type
    Blog Post
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...post Read the paper in Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control...paper in Nature Methods Find the anti-CRISPR plasmids Tagging human transcription factors with GFP...White’s lab recently deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As...

  7. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...our plasmid collection page Find more plasmids and resources for plant resources on Plant Plasmids Collection...box” for its common use in plasmid manipulation and genetics research. Plasmids are ideal tools for studying...Model Organisms blog post Read more about plasmids in our Plasmids 101 blog series Learn more about CRISPR...disease models.  Check out Addgene’s collection of plasmids containing Rattus norvegicus genes! Mexican axolotl...several vectors for use in the axolotl including plasmids for use in CRISPR and Tol2 transposase induced... and after embryogenesis. Addgene has over 600 plasmids that contain Xenopus genes and a tool-kit from...transformations and the use of Arabidopsis in the classroom on our blog. Now that you’ve learned more about...

  8. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ... were in the cytoplasm of dendrites, and only a small proportion were found on the plasma membrane (top...to gold particles localized in the cytoplasm or bound to the plasma membrane. Note that the use of the... the DREADD receptors need to be located on the plasma membrane to be activated by the DREADD actuator...membrane in mice neurons, most of them were still cytoplasmic. Use of ‘small’ tags to identify proteins Given... the limited expression of hM4Di‐mCherry at the plasma membrane in monkey neurons, we next hypothesized...might improve the transport of this DREADD to the plasma membrane. The haemagglutinin (HA) tag is a small...tag resulted in abundant expression of hM4Di in plasma membranes in both species (bottom panel in Fig....

  9. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ...service features 210 plasmids: empty backbones, fluorescent protein tags, SARS-CoV-2 plasmids, etc. It is our...to Elute and Store Your Plasmid DNA? by Paolo Colombi Should I store my plasmid in TE, Tris, or water?...Here we cover five methods to do this. 5. Plasmids 101: Plasmid Incompatibility by Leah Schiesow If you're... We've had 11 blog posts on COVID-19 featuring plasmid tools from depositors who quickly shared their ...you're thinking about hosting multiple plasmids in one cell, it might be that you can't stably maintain...This blog post features two mechanisms that cause plasmids to be incompatible. 4. RNA Extraction Without...CRISPR to detect SARS-CoV-2, including many with plasmids deposited at Addgene! 2. DNA Purification Without...

  10. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ... target a cell class in mice is not a good predictor for targeting the same cell class in NHPs (R=0.38... target a cell class in monkeys was a good predictor for targeting the same cell class in humans (R=0.67... GFP marker (CatCH-GFP). The lab deposited AAV plasmids containing the active promoters at Addgene. Performance...specific cell types (ex: rods or cones) or cell classes (ex: photoreceptors which encompassed both rods...targeting More than 70% of expression was from a single class of cells  Of the four different promoter designs...of the retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic promoters performance in ...the odds of that promoter targeting the same cell class in all three species are still greater than random...

  11. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ... in E. coli with two plasmids (Figure 3 below): Input Plasmid: The input plasmid (high copy, ColE1 origin...inducible promoter (PPhlF).  Output Plasmid: The output plasmid (single copy, bacterial artificial chromosome...through Addgene. Their plasmid maps and GenBank files can be found on their plasmid pages. All two-input...indicated figures in our paper (1)) Input Plasmid Output Plasmid Fig. 3A pNR64 pNR160 Fig. 4A pNR220...Materials and Methods section of our paper (1). Lastly, it is possible to build gene regulatory RSMs (...two-input RSMs and three three-input RSMs. The plasmids used to construct these RSMs are listed in the...paper (1). These parts are also annotated on the plasmid maps and GenBank files uploaded to Addgene. We ...

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

    Type
    Blog Post
    ... tube is mislabeled or someone grabs the wrong plasmid? What if bacteria snuck into the prep during the... human error, fragments of DNA from the helper plasmids or the cell genome can be packaged inside the ...) are aligned to the reference sequence of the plasmid used to create the viral prep. In a clean viral...this reference sequence. Second, we perform a megaBLAST search on all the reads that did not map to the...bacterial genomes, cloning vectors, and helper plasmids (Chadeuf et al., 2005, Wright et al., 2011). In...correspond to the contaminant. Each contig is then BLASTed against the NCBI nucleotide collection database...database, and it may also be aligned to plasmids in our inventory containing the suspected contaminating sequence...

  13. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...in a 1-plasmid system, in which Cas9 is included on the gRNA-containing plasmid, or a 2-plasmid system...contain thousands of gRNA plasmids, discerned only by a unique barcode on each plasmid. As such, sequencing...are composed of many different gRNA-containing plasmids, each with a different genomic target. The goal...sequences and ease of generating gRNA-containing plasmids. CRISPR genome wide-screening experiments commonly... target every gene in the genome, or a specific class of genes? Addgene currently carries several genome-wide... selection of sub-libraries targeting specific classes of human genes. What are the steps involved in ...other types of experiments, check out our CRISPR Plasmids and Resources Page. References McDade JR, Waxmonsky...

  14. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...Baird, who has created plasmids as a technician in the Davidson lab for the last four years. “Researchers...Molecular Expressions collection including some 3,300 plasmids along with image galleries and educational resources...resources to go with them. This collection of plasmids is available in an easily searchable format on Addgene...Fluorescent Protein Collection webpage. These plasmids are divided into the following categories: Over...list of cell markers all tagged with mEmerald. Plasmids to monitor protein-protein interactions through...Specialty constructs for biosensing. Over 300 plasmid backbones spanning a rainbow of fluorescent proteins...Proteins - Making the Right Connection.  mEmerald plasmids and many more! Baird has a helpful tip for anyone...

  15. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...bacteria with the plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows... Inside Addgene Posts! Learn all about Plasmids in our Plasmids 101 series! ...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the...about Developing Lab Management Software    The plasmid's journey begins First, each package is opened and... to hundreds of samples, with an average of 10 plasmids per deposit. Each week an average of 16 deposits...allow us to store multiple types of tube for each plasmid we have at Addgene. Each sample at Addgene has ...strains that have been treated to easily take in plasmids (transformation). Transformed bacteria are then...

  16. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    ...Would you buy and use a plasmid if it didn’t include any plasmid sequence data? Of course not. Unfortunately...depositing your plasmids encoding antibodies, ScFvs, and nanobodies to Addgene! The plasmids will be made...Plückthun, 2015a, b). Recombinant antibodies are plasmid-based and offer numerous advantages, including ...antibodies that you have come to expect from our plasmid and viral services. The first step will be working...working with the community to build a resource of plasmids encoding recombinant antibodies as well as other...through our usual service (see our current antibody plasmid collection) and we plan to introduce affinity reagents...through distributions of research materials, such as plasmids and viral vectors. However, it typically takes...

  17. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...For the last 14 years, scientists have been coming to the Addgene website in search of plasmids. Now, scientists...synthetic biology page Search for a plasmid Find Addgene's most popular plasmids ...Addgene as a large data set. Addgene has over 65,000 plasmids in the repository, each verified by sequencing...at its sequence. MIT collaboration for gene classification This inquiry started when Neil Thompson, a ...is synthetic or natural with 97.7% accuracy. Classifying genes as natural or synthetic But how can you...organism, it may express protein more efficiently. To classify gene sequences as natural or synthetic, the researchers...simulations and machine learning, they created a classifier that predicts whether a gene was natural or synthetic...

  18. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...single plasmid transformation, though, a pooled library has hundreds to millions of distinct plasmids, all...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...amplify the library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA,...own protocol that amplifies without disrupting plasmid representation. A good place to start is with a...

  19. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...that the plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe... 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK... Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b) Methods to ...overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T ...mammalian species to produce gas vesicles. New mARG plasmids Our new mARGs come as a set of two PiggyBac integratable...genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient transfection... into the genome by cotransfecting them with a plasmid expressing PiggyBac transposase. In rtTA-cell lines...

  20. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ....org Find Cre-Lox Plasmids Find Optogenetics Plasmids Find Chemogenetics Plasmids ...vectors expressing light inducible ion channels (plasmid # 20298 for example) have revolutionized Neuroscience.... These Cre/ FLP versions are retained in the cytoplasm until estrogen receptor ligand Tamoxifen is added...neurons in the living brain   Figure 1: Plasmid mix to label neuronal morphology (eYFP) and the...recombinase. After in-utero electroporation, this plasmid mix results in sparse labeling of cortical neurons...labeling of individual neurons one could reduce the plasmids amounts to reduce transfection efficiency. However...visualized at the same time from two different plasmids this approach becomes problematic as it reduces...
Showing: 261 - 280 of 1038 results