We narrowed to 803 results for: Cre
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TypeBlog PostPublishedApril 25, 2019, 3:01 p.m....University of Alberta and University of Oregon created mTFP1 by transforming E. coli with a gene library...tetramer-breaking mutations. The library was then screened through successive rounds of mutagenesis for the...filter. This combination results in a 2.6-fold increase in signal relative to mCerulean imaged with a ...
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CRISPR Challenges: Standardization and Homology Directed Repair
TypeBlog PostPublishedFeb. 14, 2018, 2:36 p.m....single-strand DNA incorporation (ssDI). "Easi-CRISPR for creating knock-in and conditional knockout mouse models...Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in mouse models with CRISPR has been challenging...efficiency. Easi-CRISPR uses a long ssDNA template to increase efficiency of insertion and gene replacement. ... -
In Vivo Biotinylation of Bacterial Fusion Proteins
TypeBlog PostPublishedJan. 25, 2018, 2:09 p.m....we found that adding a TEV linker dramatically increased accessibility of the AviTag and HisTag, which ...fusion protein in bacteria. Our initial attempt to create a plasmid based on the biotin ligase BirA backbone... inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available ... -
Plasmids 101: Origin of Replication
TypeBlog PostPublishedNov. 11, 2020, 2:23 p.m....R6K, and 15A. Not all origins of replication are created equal. Some will produce many plasmid copies and...because they will compete for the same machinery, creating an unstable and unpredictable environment. As ...plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain Most E.... -
Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC
TypeBlog PostPublishedSept. 21, 2023, 1:15 p.m....primary antibody and endogenous IgGs in the tissue. Created with biorender.com. Strategies for avoiding...Some labs have found that you can significantly decrease background staining by mixing your primary and...you add your primary and secondary antibodies. Created with biorender.com. Proper tissue processing... -
Antibodies 101: Chimeric Antibodies
TypeBlog PostPublishedApril 25, 2023, 1:15 p.m....Recombinant antibodies are plasmid-based and are created synthetically or derived from hybridoma antibody...antibodies against the new Fc region. In addition to increasing multiplexing capabilities this process is frequently...Although isotype conversion is routinely used to increase the utility of recombinant antibody tools, the... -
Summer SciComm: Science Storytelling
TypeBlog PostPublishedSept. 16, 2022, 1:15 p.m.... - not just for fiction! Image credit: Kenneth Rowe, under the creative commons license. The introduction...is to find one or two roadblocks that sparked creativity, changed the outcome of the project, or illustrates... -
Addgene Begins Distribution of Recombinant Antibodies
TypeBlog PostPublishedMarch 28, 2022, 4:30 p.m....collection of antibodies available at launch were created by James Trimmer, PhD, of the University of California...commitment to supporting your research, Addgene is creating an array of educational materials to accompany...more useful to everyone over time. Through our incredible community, including your support, we have enabled... -
RUBY-Red Siliques
TypeBlog PostPublishedMarch 1, 2022, 2:15 p.m....to select against CRISPR positive plants, but screening these can be challenging and requires specialized...Willoughby. Using RUBY as a selection marker The creators of RUBY highlight its use in crop plants under...as positive selection of transformed plants to decrease the routine usage of antibiotics which may contribute... -
Plasmids 101: TOPO Cloning
TypeBlog PostPublishedOct. 27, 2016, 2:30 p.m....cassette allowing a researcher to perform blue-white screening after transformation - self joining of the vector...break down the steps needed for TOPO cloning: 1. Create Your PCR Product: Design standard primers (no need...unpaired 3’ ends in your PCR product. If you need to decrease error rate, please use one of these methods to... -
Don’t Wait To Launch Your Health Science Startup
TypeBlog PostPublishedAug. 25, 2015, 2:30 p.m....would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am lucky... your potential as an entrepreneur, not your credentials. If you have an idea, don’t wait. Go for it.... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog PostPublishedMarch 14, 2019, 1:54 p.m....microvesicles (approx. 40 - 150 nm diameter) that are secreted from the cellular membrane into the culture medium...AAVs (Schiller et al., 2018). Exosomal vesicles secreted from producer cells also contain cellular macromolecules...open new ways of improving gene delivery. They increase the efficiency of transduction, broaden the use... -
Virtual Science Conference Coming Up? Three Addgenies Share Their Experience
TypeBlog PostPublishedJune 30, 2020, 1:15 p.m....to keep the audience engaged through a computer screen. For the Reproducibility for Everyone Workshop,...author of the poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different...canceled may be disappointing, but there are many creative ways to adapt and take advantage of the new conference... -
Four Factors that Differentiate the Stem Cell Field
TypeBlog PostPublishedSept. 28, 2016, 2:35 p.m.... the stem cell community – a community largely created due to Yamanaka’s seminal work. It didn't take... during his keynote address, appeared almost incredulous that the field has exploded so quickly, citing...explained that it took 50 people nearly 2 years to create the first stem cell line that could be used in ... -
Plasmids 101: Mammalian Vectors
TypeBlog PostPublishedMarch 25, 2014, 3:15 p.m...., described below, are typically employed when creating stable cell lines and are not used for general...Stable transfection A stable transfection is used to create a population of cells that have fully and successfully...culture media until 24-48 h post transfection when creating stable cell lines. Looking for Mammalian Expression... -
Behind the scenes: Addgene’s new search engine and more
TypeBlog PostPublishedDec. 12, 2019, 2:22 p.m....well-characterized. During this time, we’ve also created a variety of educational resources in an effort...understanding of Addgene’s customer’s needs. Categories We created a categorization of our information in order to...for promoter and serotype, among others. We also created a normalization mechanism to “collapse” variations... -
15 Years of Addgene: The Top 15 Plasmids
TypeBlog PostPublishedJan. 8, 2019, 1:54 p.m....sequence (pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP ...virus, making it ideal for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked...site, a pair of annealed oligos can be cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9... -
Luminescent Imaging with Nano-lanterns
TypeBlog PostPublishedMay 25, 2017, 2:30 p.m....emitted yellow-green light and exhibited 3-5 fold increased brightness over previous BRET probes, with sufficient...injected intravenously with coelenterazine, with increased sensitivity and faster imaging compared to previous... reporter genes for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent ... -
Addgene’s Viral Service - Why Virus? Why Now?
TypeBlog PostPublishedMarch 23, 2017, 2:30 p.m....production. Finally, we are blessed to have an incredibly supportive Board of Directors at Addgene. Our...leadership of Addgene’s co-founder, Melina Fan, we created a new team to push this project forward. The viral...you to the Addgenies who went the extra mile to create this new service. We knew the scientists were waiting... -
Communicating Your Science With Help From ComSciCon
TypeBlog PostPublishedJuly 19, 2018, 1:12 p.m....communication skills with training programs, which are increasingly available. But think of this as a jumpstart ...possible outcome of a ComSciCon workshop is the creation of a new science communication initiative that...science communication opportunities in your area, create them. Seek out collaborations with organizations...