We narrowed to 816 results for: LAS
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TypeBlog PostPublishedJuly 28, 2016, 2:30 p.m.... 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 ...
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AAV Vector Quality Control: Going the Extra Mile with NGS
TypeBlog PostPublishedSept. 12, 2017, 1:44 p.m.... 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... -
Genome-wide Screening Using CRISPR
TypeBlog PostPublishedAug. 18, 2015, 2:30 p.m....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... -
The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins
TypeBlog PostPublishedJuly 29, 2014, 6:16 p.m....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... -
Tips from the Repository Trenches: Using Barcodes to Track Samples
TypeBlog PostPublishedOct. 6, 2015, 2:30 p.m....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... -
Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource
TypeBlog PostPublishedMay 25, 2021, 2:29 p.m....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... -
Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation
TypeBlog PostPublishedNov. 8, 2018, 3:47 p.m....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... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 19, 2017, 1:54 p.m.....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... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog PostPublishedJuly 13, 2023, 1:15 p.m....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... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog PostPublishedJan. 24, 2023, 2:15 p.m....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...