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We narrowed to 812 results for: Cre

Showing: 451 - 500 of 812 results
  1. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ...individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library. The fixed...detected over a 10-week period. Their expression decreased over time but they were not silenced.    Find ...retroviral overexpression cocktail yielded a three-fold increase of fully reprogrammed cells based on single-cell...
  2. Plasmids 101: Origin of Replication

    Type
    Blog Post
    Published
    Nov. 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....
  3. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 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 ...
  4. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 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. ...
  5. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    Published
    Jan. 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 ...
  6. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 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...
  7. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    Published
    Aug. 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....
  8. Virtual Science Conference Coming Up? Three Addgenies Share Their Experience

    Type
    Blog Post
    Published
    June 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...
  9. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 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...
  10. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    Published
    Sept. 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 ...
  11. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 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...
  12. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    Published
    Feb. 8, 2022, 2:15 p.m.
    ...use small, incremental changes to understand what works and what doesn't. By being creative, thoughtful...each step as you complete it. Yes, this can be incredibly tedious, but it also works, so do it anyway.  ...
  13. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 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 ...
  14. Addgene’s Viral Service - Why Virus? Why Now?

    Type
    Blog Post
    Published
    March 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...
  15. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 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...
  16. Communicating Your Science With Help From ComSciCon

    Type
    Blog Post
    Published
    July 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...
  17. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    Published
    July 22, 2025, 1:15 p.m.
    ...reaction catalyzed by luciferase, releasing light. Created with BioRender.com. Bioluminescent reporters...Peak emission (nm) Size (kDa) Substrate Cofactors Secreted North American firefly (Photinus pyralis) Natural...cell membrane and deliver the substrate; however, secreted luciferase or alternative substrates can permit...
  18. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ... vector and insert are (separately) digested to create 10–12 base “chewed back” overhangs, they can be...bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened up a...transformed into bacteria, which repair the plasmid. Created with BioRender.com. How does SLIC compare to...
  19. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ...under cold temperatures, and frigid conditions decrease the potential for microbial contamination. Additionally...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate cold-tolerance...The event also featured a seminar on “Tools to increase scientific reproducibility” by Benoit Giquel, ...
  20. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    Published
    Dec. 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...
  21. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    Published
    Feb. 11, 2021, 2:15 p.m.
    ...and their GPCR binding partner, Bryan Roth’s lab created an open source resource called PRESTO-TANGO (parallel...parallel receptorome expression and screening via transcriptional output - TANGO) (Kroeze et al., 2015...are ideal candidates for use in chemogenetics. To create chemogenetics receptors, GPCRs were first mutated...
  22. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...mutations through homologous recombination. To further increase the utility of CRISPR/Cas9, it will be necessary...that enhance pol III transcription, potentially increasing the levels of gRNA expression. Since tRNAs are... of multiple gRNAs targeting a single gene can increase the likelihood of gene knockout, or multiple gRNAs...
  23. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...funding and time. While institutions that focus on creative problem solving like the Howard Hughes Medical...plant research, more initiatives are needed to increase the body of knowledge. Organizations like Open...research and effectively carry out their mission of creating new forums and opportunities for open innovation...
  24. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    Published
    May 22, 2014, 4:02 p.m.
    ... the world of science publishing, and an ever increasing number of journals and publishers to choose from...them needing a subscription to the journal. An increasing number of funding bodies insist that the research...is beneficial for both the referees (as they get credit for their time and effort) and the scientific community...
  25. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ...publishing has a number of drawbacks, especially the increasingly growing time from submission to publication....for their efforts, inspire future research, and increase their impact. Open access and transparency Another...journal publication or an updated preprint, further increasing their findability. Now, go out there and finish...
  26. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    Published
    Oct. 29, 2019, 2:52 p.m.
    ...electric! After attending this meeting I had an increased appreciation for the role of ion channels and ... the entire worm during regeneration, the worm creates two heads. Subsequent cutting results in a two-...pHLuorin fluorescent protein. This GEVI features increased brightness and improved plasma membrane specificity...
  27. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    Published
    Oct. 1, 2019, 1:21 p.m.
    ... viewed your profile.”  The latter is much less creepy. If we’ve met before, I’d be much more likely to...borders on my first rule of networking--”If it feels creepy, don’t do it.” It’s just not a useful way to build...rules of good resume writing. Include examples of concrete accomplishments in both the technical and “plus...
  28. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    Published
    Jan. 31, 2019, 1:32 p.m.
    ...cells in the microenvironment. This results in increased expression of transforming growth factor-β signaling...signaling, possibly inducing the secretion of growth promoting proteins, such as human growth factor, vascular... (Kogure et al., 2011) believes that miRNA may decrease the expression of TAK 1, an enzyme that controls...
  29. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...figure out how to optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later,...reagents.” Here at Addgene, we’re excited to see the creative new ways you put the ClonTracer library to use... Blog Learn about the Sabatini and Wood Drug Screening Kit Find Out How to Best Utilize Our Cancer Resource...
  30. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...available on our FRET Resource page and can be used to create fusions with a gene of interest. These plasmids...biosensors too, as another laboratory may have already created the sensor that you need. FRET biosensors designed...multiple FRET biosensors simultaneously would further increase the amount of correlated information available...
  31. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...this through a variety of activities, including creating educational materials and lesson plans that are...sterilization of women in marginalized communities have created a legacy of distrust between these communities ...communities, including those by researchers hoping to increase community participation in new precision medicine...
  32. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...response to blue light. This tool, CRY2clust, was created by adding a 9-residue peptide to the C-terminus...regions. To overcome this issue, Shemesh et. al. created a soma-targeted derivative of the opsin CoChR, ...within the cell. Their Fluorescence fLuctuation INcrease by Contact (FLINC)-based biosensors measure fluctuations...
  33. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...  Neurogenetic alchemy: CAP-Mac unleashes the secrets of primate brains with brilliance By Hridaya Patel...authors used a multispecies approach to generate and screen a library of AAV9 variants (Chuapoco et al., 2023... Figure 3: Overview of library generation and screening that led to identification and characterization...
  34. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...Figure 1: Common functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids...resistance genes to help cells survive their own secretions. Pro tip! Historically, colicin-producing plasmids...years, and new classifications are still being created as whole-genome sequencing becomes more relevant...
  35. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    Published
    Jan. 9, 2020, 1:45 p.m.
    ...growth or decline of interest in a field over time. Creating an alert for a search term helps automate your...is not a traditional search engine. Instead, you create your own research feed by entering keywords, concepts...new papers that contain these terms. You can also create intersects in your feed which means only papers...
  36. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...activation or repression, transgenic mouse line creation, and in vivo gene editing of mice, with the potential...cationic polymer. Polybrene has long been used to increase retroviral transduction efficiency by neutralizing...Nanoblades CRISPR has also been extensively used to create transgenic mice by microinjecting zygotes with ...
  37. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...using the plexWell technology to easily and quickly create Illumina sequencing libraries. From start to finish...ask the depositing laboratory to review the discrepancies. If the depositing laboratory confirms that ...-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863 . https://doi.org/10.1038...
  38. 6 Tips for Grant Writing

    Type
    Blog Post
    Published
    Oct. 18, 2016, 2:30 p.m.
    ...tips to improve your grant writing and hopefully increase your success rate. 1. Submit your grant early...are unlikely to get past the initial round of screening. I STRONGLY recommend that you ask a PI or mentor...taxes or charitable donations) so there is an increasing realization that, if we want people to keep contributing...
  39. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...Mammalian expression plasmids are primarily used to create mRNA and the commonly used mammalian terminators...below) releases the mRNA from the polymerase and creates a free 3' end which is now available for polyadenylation...Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered Transgenes...
  40. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    ...Victoria, CA Over the decades science has become increasingly restricted to academic and industrial labs, ...research. This interest manifested itself in the creation of Hackerspaces and Makerspaces back in 2007, ... make accounts with  Addgene. We now have the incredible CRISPR/cas9 tools at our disposal to to modify...
  41. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...parallel because the risk of accidental errors increases. In addition to human error, fragments of DNA ...to the reference sequence of the plasmid used to create the viral prep. In a clean viral prep, like AAV...control process! The images in this blog post were created using Snapgene and Geneious software. References...
  42. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...disulfide bridges, glycosylation and acetylation to create functional, stable proteins. Protein expression...scientists have taken advantage of the natural secretion of biochemicals and proteins through plant roots... Tagging recombinant proteins with a naturally secreted plant peptide allows for easier access and purification...
  43. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used...
  44. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    .... At Addgene, we use blastn to identify any discrepancies in Sanger sequences, such as mismatches, deletions...determine the potential effect of any nucleotide discrepancies. The blastp and blastx programs are optimized...plasmids instead! All BLAST images are modified screen shots from the NCBI BLAST website. Additional Resources...
  45. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...expense and requirement for lab equipment. Plus they create plastic waste. Kits can also have the annoying ...and filters can be stacked together in series to increase binding capacity. Figure 1: Purify DNA using...Additionally, while commercial plasmid prep kits come in discreet sizes (mini, midi, maxi, giga) which requires ...
  46. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...a double-edged sword. An antibody’s large size creates more distance between the target antigen and the...also allows for better tissue penetration and decreases the distance between a fluorescent tag and the...HCab antibodies from its serum, and meticulously screening, optimizing, and characterizing several anti-mouse...
  47. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores, compromising the photon...improved fluorophores are making chemical dyes increasingly attractive and accessible to cell biologists—and...fluorogenicity, photoactivation etc.—can further increase the photon budget and allow us to push further...
  48. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    Published
    Nov. 14, 2017, 2 p.m.
    ...just might personally hand your credentials to HR, greatly increasing your chances of getting a phone ...can use to better prepare for your career, and increase your chances of landing that next coveted job!...
Showing: 451 - 500 of 812 results