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We narrowed to 799 results for: Des

Showing: 241 - 260 of 799 results

  1. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...Cas13a as a diagnostic tool. A pool of DNA or RNA nucleotides containing a sequence of interest (red) is amplified...Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex with crRNA...the target sequence is present in the pool of nucleotides, the nonspecific RNAse activity of Cas13a becomes... sequence is present in the original pool of nucleotides.   What are the potential applications of...can mediate cleavage with spacers from 20-28 nucleotides in length and does not require a specific PFS... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display...

  2. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ...scarless mutations, ones that don’t leave any undesired scars in the chromosome. These methods roughly... recombination enzymes can also accomplish the desired genomic modification. This technique is frequently...resolution step (“double crossover”) which leads to the desired mutation and excision of the vector backbone. This...the pDS132 MCS with a versatile, synthetic MCS (designed with the help of the DNA tuner program) (Latynski...purified amplicons upstream and downstream of the desired deletion directly into cut vector with excellent... of interest, from the day primers arrive, the desired chromosomal modification can be made in 5 days....

  3. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool for understanding...organisms lacked a classic cysteinyl tRNA synthetase, despite abundant incorporation of cysteine in their proteomes...pSer-incorporated proteins on a single plasmid. This includes 4 copies of a tRNA that specifically recognizes...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...

  4. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    Published
    Sept. 1, 2015, 2:30 p.m.
    ...Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc. “How much do you need [for your...necessary resources. This is why at Experimental Designs Consulting (EDC), we instead have our clients focus...the institution/funding source. Major equipment includes large centrifuges, sequencers and analyzers, and... as many of these tools as possible, this even includes clocks, timers, and phones.  As for reagents and...passion for science. Follow him on Twitter @XpDesignsMGNT        Additional Resources Find other career...the Addgene Blog! Learn more about Experimental Designs Consulting at their website. ...

  5. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...R26R-Confetti described in Hugo J. Snippert, et al. (2010) and the MAGIC Marker strategy described in Karine...Brainbow-1 system employs Cre-mediated deletion, the design of Brainbow-2.1 cleverly combines Cre-mediated ...protein sequences are highly homologous, preventing design of antibodies specific to each fluorophore. Third...immediately following the promoter. The STOP cassette includes a mutant YFP that does not fluoresce, but can ...produces a minimum of 8 colors. The use of AAV provides spatial and temporal control without the need ...

  6. Newly Updated AAV Data Hub!

    Type
    Blog Post
    Published
    Feb. 7, 2023, 2:15 p.m.
    ...We're excited to roll out the redesigned, revamped, and re-launched Addgene AAV Data Hub, the place [email protected] to learn more. A great submission includes: Images that clearly show viral spread and cellular...your experiment* Functional data, if the report describes a functional vector such as a channelrhodopsin...site as well as the target region, if the report describes a retrograde serotype An abstract that summarizes...work. Sharing your negative data helps others to design better experiments (and wouldn't it be nice if ...

  7. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

    Type
    Blog Post
    Published
    Nov. 12, 2014, 3:54 p.m.
    ...researchers at Japan’s Hiroshima University who described their new tool in Scientific Reports in June. ... gap. As Sakuma and colleague Takashi Yamamoto describe in their recent study, the construction system...vector-mediated multiplex genome engineering. The system now provides an efficient targeting strategy for multiplex ... validated and ready. “Currently, the kit only includes two versions of Cas9, nuclease and nickase, but...Addgene here. Other Popular CRISPR Posts: How to Design Your gRNA for CRISPR Genome Editing CRISPR-Cas9...

  8. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ...antibody worked under the conditions described above, and a short description of how the antibody performed....the submitted application, along with a brief description of the experiment. If that data was included ...ELISA — but in all cases, it contains a detailed description of the experimental conditions.     Figure 4...The submission process is easy, with simple, descriptive fields, drop-down boxes when appropriate, and...

  9. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 2:15 p.m.
    ... capsid of another. Formally, pseudotypes are described with two numbers separated by a slash: the first...cardiac, and retinal cells (Issa et al., 2023). AAV2 Despite being isolated second, AAV2 is the best studied... choice of AAV vector inherently depends on the design of your experiment. Though we cannot make universal...experimental reports on commonly used vector cargo designs and administration doses. Don't forget to submit...2008). Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1,...., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV Variant Permits Efficient Retrograde Access...

  10. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...In 2005, Boyden et al. described the first light-activated tool for controlling neuronal activity, channelrhodopsin... light in combination with retinal) variations designed to give researchers flexibility in using optogenetic...precise timing of the perturbation. Other tools are designed to be activated within a narrower band of wavelengths...function through multiple intra-cellular signaling cascades. Optical methods for controlling GPCR activation...control GPCR activation and downstream signaling cascades (reviewed in Abreu & Levitz, 2020), including ...You’ll need to test for these differences when designing your experiment.  Finding the right information...

  11. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...colleagues describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six... bring this into your target organism. We have designed a Golden Gate system based on the daily needs ...and cut another,” Lohmann said. “It allows us to design modules that always have the same recognition site...produced will have different overhangs, they can be designed such that they go back together only one way and...

  12. Editor's Choice, September 2016

    Type
    Blog Post
    Published
    Oct. 7, 2016, 4 p.m.
    ...record breaking month and to find other posts that deserve a second look. 5 Great Apps for Lab Life This very...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ...Jessica Sacher from the University of Alberta and describes a technique you can use to find phage proteins...molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique to screen for...that, while affecting a small number of people, deserves our attention and has huge potential for the development...

  13. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    Published
    March 22, 2016, 2:30 p.m.
    ...so he assembled a plasmid kit designed to do exactly that. As described in the journal Science Signaling...When cancers are treated with drugs designed to hit them right where it hurts, the effects are often ...mutant complementary DNAs (cDNAs) representing core nodes that, when over-expressed, constitutively activated...activators for more cancer-related pathways and more nodes within those pathways. With support from the National...

  14. SciComm with the Experts at Science in the News Part 2

    Type
    Blog Post
    Published
    April 27, 2017, 2:30 p.m.
    ... discusses some of the programs SITN runs and describes some of the more popular topics from the Science... fields as well as students from the ed school, design school, and Kennedy school to discuss the impacts...impacts of science in a social context. Episodes are driven by narrative and story telling. Chapter 5: The...The SITN Blog - Vini and Amy describe how the SITN blog is a source of scientific information for the ...underwater? Chapter 8: Work-Life Balance - Vini describes how she balances working in the lab and working...

  15. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    Published
    May 17, 2016, 2:30 p.m.
    ... be spent making and testing entirely new gRNA designs. You can now easily find many validated gRNAs in...validated gRNAs? Proper target selection and gRNA design are essential for a CRISPR experiment.  The selection...headers to sort alphabetically or in ascending or descending order, in the case of numeric columns.  The data...Additional Resources on the Addgene Blog Learn How to Design Your gRNA Use the CRISPR Software Matchmaker Browse...Additional Resources on Addgene.org Browse gRNA Design Tools Read Our CRISPR Guide Pages Find CRISPR Plasmids...

  16. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    Published
    July 9, 2015, 2:30 p.m.
    ...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...

  17. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ... antibody sequences, degenerate primers can be designed to selectively mutate specific regions of your...sections. Structure-guided directed evolution If you desire a bit more control, structure guided directed evolution...complex as a template. This structural framework then guides the specific mutations you choose, whether it's...allows you to combine the strengths of rational design with the power of directed evolution, all while...technologies, the Institute of Protein Innovation (IPI) provides information on their large-scale recombinant antibody...synthetic antibody products and development methods. Despite recent advances, there is still always room to ...

  18. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ... sequence that moves around. This DNA sequence encodes the proteins the transposon needs for transposition...contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase and integrase enzymes...ORFs) which often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities...preferred target site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from...encode reporter genes, mutagenic cassettes, or barcodes. When transposons are delivered to cells or a ...occurring Tn7 transposon found in Vibrio cholerae that encodes a Type I-F CRISPR-Cas system. This system has four...

  19. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...eukaryotic process where approximately 20-30 nucleotides of double-stranded RNA (dsRNA) results in downregulation... of an RNAi-inducing transgene. Typically, the desired target of RNAi is expressed as inverted repeats... and quantify actively silencing plants. Non-destructive methods, most popularly with a fluorescent reporter...Fluorescent reporter-based RNAi constructs have been designed and widely implemented in plants, such as in the...approach, myself and others in the Luis Vidali lab designed a new RNAi method. Instead of using a fluorescent...functional APT will convert 2-FA to cytotoxic nucleotides, resulting in death. In the presence of 2-FA,...

  20. Antibodies 101: Affinity Tags

    Type
    Blog Post
    Published
    Sept. 19, 2023, 1 p.m.
    ...You’ve designed the perfect experiment – controls, conditions, and everything in between – now all you...This is where affinity tags come in – they are peptides which do not naturally exist in most expression...protein for downstream applications Below, we describe a few of the most common tags available. Maltose...antibody is an alternative to amylose resin and also provides a visual detection mechanism via applications ...especially if downstream antibody detection is desired… and yes, we have an antibody for that!   ...
Showing: 241 - 260 of 799 results