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We narrowed to 929 results for: Comp;

Showing: 501 - 550 of 929 results

  1. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...For example, Drosophila lines with compound autosomes are completely infertile when mated to wild fruit...with appropriate measures. Barriers should be a component of all gene drive confinement strategies, but ...: I. Cage Studies of Chromosome Replacement by Compound Autosomes in Drosophila melanogaster. Genetics...

  2. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ... an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused to an RNA adenosine...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

  3. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...orders were affected by the COVID-19 pandemic, we compared the number of plasmid orders and new COVID-19 ...the likelihood in our Poisson process into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential...effects fairly well. However, we observed one complication in the China data – there is a period of 1 month... lockdown in China, then the observed data is compatible with an infinity-fold reduction in activity due...

  4. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...real time. When the PCR is complete, the results are analyzed by comparing the threshold fluorescence ...amplification reaction to take place.  When the PCR is complete, a droplet reader extracts the droplets from the...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...

  5. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    ...this situation: You’ve spent hours scouring a company’s LinkedIn pages, given yourself the bathroom mirror...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...the candidate has what it takes to thrive at the company and add value, not just “do the job”. When they...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...

  6. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...gross comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids....PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived from. They also...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

  7. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...them to near homogeneity using a 6XHis tag, and compared their specific activities by measuring NADH production...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...

  8. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...system, the possibility of creating a replication-competent virus via recombination between the transfer and...recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition...Lentiviral barcoding libraries have been used for complex applications like lineage tracing in heterogeneous...

  9. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money... few ways you can do this. If you are going to complete your colony PCR analysis in the same day, you ... product size on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel... larger sized products for positive clones (+) compared to negative clones. Finally, orientation-specific...

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

    Type
    Blog Post
    ...is ideal for tracking large collections. Many companies, such as Thermo Fisher and Micronic, provide 2D... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...specific barcodes etched or printed onto them by companies such as Tubewriter. Any lab can benefit from using...tubes themselves. Although we’ve developed our own computer system for tracking samples, it can be done easily...

  11. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...survive as a researcher because of the tough competition for academic positions and poor grant situation...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...

  12. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...on OWW, including those for preparing your own competent cells. If you’re familiar with the process of ...service free of charge to scientists by allowing companies to pay publish their catalogs on the site. Like...of checking off each step as you painstakingly complete your experiment – one small victory in a long ...manage scientific data: Electronic lab notebooks compared with Google Docs and wikis - by Rory Macneil on...

  13. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...target provided that (1) the sequence is unique compared to the rest of the genome and (2) the sequence...for the canonical S. pyogenes PAM sequence NGG compared to wild-type SpCas9, which also displays some ...CjCas9) is even smaller than SaCas9 and is also compatible with AAV delivery. NmCas9, another small ortholog...(2018) Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 ....

  14. Hot Plasmids - August 2020

    Type
    Blog Post
    ...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate...

  15. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn...variants that reached the CNS following IV injection. Compared to AAV9, PHP.B has a >40-fold increase in CNS ...rat CNS: AAV PHP. EB vector dose–response and comparison to AAV PHP. B." Gene therapy 25.5 (2018): 392...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

  16. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...are needed to deliver all the protein and RNA components, and several sgRNAs must be screened to identify...eBook Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

  17. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ... and the complexity of effectively deploying them in the lab somewhat mirrors the complexity of understanding.... Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both to..., and jGCaMP8 viral preps distributed per year compared to other AAVs preps from 2016–2023. B) Inset showing...

  18. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...virus as self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...virus’s most abundant proteins and a critical component for viral genome packaging. Addgene recommends...

  19. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...in order to be expressed stably in neurons. As compared to epitopes tags, FPs are intrinsically fluorescent...fluorescent proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags. Indeed,... structure resembles images of the deity (see comparison image above). The church of the Flying Spaghetti...

  20. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...

  21. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...meets two conditions:  1) The sequence is unique compared to the rest of the genome.  2) The target is present...found. If you’re lucky enough to find a SpCas9 compatible PAM at your site of interest, that is the most...expanding number of Cas9s with looser PAM requirements compared to SpCas9. xCas9 has a PAM requirement of NG, ... proteins. The Synergistic Activation Mediator complex, generally the strongest activator, consists of...

  22. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Host range depends on whether the host has the components to recognize the ori and allow for replication...organisms. pBBR1-based plasmids and plasmids in incompatibility groups N, P, Q, and W are known to have replicons...list of the bacterial species these plasmids are compatible with.  Table 1: Non-exhaustive list of common... and Srivastava 2013. Family Replicon Compatible with Shown to work in IncN pCU1 Gram-negative...

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

    Type
    Blog Post
    ...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...

  24. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136...Kadoch C (2018) Interrogation of Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale...

  25. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...endogenous homologous recombination enzymes can also accomplish the desired genomic modification. This technique... which results in a merodiploid. This can be accomplished by selecting for an antibiotic resistance marker...scarless modification of bacterial genomes. It can accomplish large deletions, insertions, and even precise...PMC4513841. Latynski US, Valentovich LN. "DNA tuner: a computer program for the construction of polylinker sequences...

  26. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ... proteins in living cells and organisms. Both components of the chimera are encoded by DNA. Since researchers...When both orientations are available, a direct comparison of these fusion proteins may reveal which of ...the fusion protein can be verified and it can be compared to what is expected based on immunofluorescence...interactions) can be challenging. If a fusion can complement a knock-down or knock-out of the corresponding...

  27. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...the production of lentiviral vectors that are competent for one round of infection (AKA transduction in...progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with...double-stranded DNA transgene is incorporated into a large complex of both viral and cellular proteins and trafficked...titering IDLVs include detecting vector particle components or performing quantitative PCR titration of vector...

  28. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...genetically encoded tag and exogenously added components. For a review see Lotze et al. (2016). To date...is used to excite GFP, may also excite several components that are naturally present in cells or media....

  29. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ... (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv.   What...their lack of an Fc region. Disadvantages scFVs:  Compared to antibodies, scFvs tend to have lower affinities...intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged with ....0000000000000145  Gąciarz A, Ruddock LW (2017) Complementarity determining regions and frameworks contribute...

  30. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...method for knockouts is Tracking of Indels by Decomposition (TIDE) (Brinkman et al., NAR). This technique...trace files of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits...This method requires a set of control cells to compare the sequencing reads from your edited sample to...assessment of genome editing by sequence trace decomposition. (2014) Nucleic Acids Research Volume 42, Issue...

  31. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...lacY1 proA2 rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning and storage of unstable ...XL10 Gold Tetracycline and Chloramphenicol High competency cloning and propagation of large plasmids, ligated...tables are by no means exhaustive! For a more comprehensive list and additional information, visit OpenWetWare's...of Bacterial Expression Systems. Check out our companion post describing protein expression strains, where...

  32. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...helps all viewers, especially if you have some complex patterns with several overlapping targets to interpret...you add the secondary antibody, it should get completely washed away and leave no signal. If you see lots...samples where you label only one target in each. Compare these individually-labeled samples with the results...deep phenotyping and spatial analysis of cells in complex tissues. Proceedings of the National Academy of...

  33. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...certain Cas enzymes are less compatible with the bulkiness that accompanies mispairing of bases. To reduce...unnecessary for many applications, it is the only comprehensive method available. Targeted sequencing approaches...must be quantified slightly differently. For a comprehensive approach, RNA-seq would be an appropriate method...

  34. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...could avoid potential off target artifacts by comparing the phenotypes of multiple gRNAs. Targeting different...lanes 0.3 % 3.0, Figure 5, lane 2) is faint, so comparing it to the undigested control (Figure 4, lane 0...instead of agarose gives much sharper bands. I compared two MOIs of gRNA transduction to see if that had...testament to how far genome engineering has come. For comparison, in 2007 it took about 3 months to knockout a...

  35. Addgene’s First AAV Data Hub Challenge

    Type
    Blog Post
    ...your data by Nov. 12 to compete. The top ten participants will be invited to compete in our speed round data.... Bobae An will host a Q&A session after the competition, where you can get all your burning AAV questions...

  36. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...In complex metazoans, rapid cell division and large scale cell mobility are essential processes during... required for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...organism. Studying cell mobility within these complex environments has never been easy and, for many ...labelling of cells using cre-lox is advantageous when compared to photoactivable or photoconvertible proteins...

  37. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...straightforward to learn and relatively cheap (compared to many other technologies and techniques in the...experiment work? What are the various tools and components necessary to run it? And, most importantly, what...writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow...applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nature methods....

  38. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...fragments, allowing ordered assembly of multiple components into a destination vector. The first step in ...Oligonucleotides specifying the gRNA seed sequence should be compatible with type IIS enzyme BfuAI. Gateway cloning is...supply it with another plasmid.   Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA...Cas9-containing binary vectors. The system is compatible with both monocot and dicot plants. Liu Lab Golden...

  39. Editor's Choice, July 2016

    Type
    Blog Post
    ...easier to generate mouse models.   Cpf1 Update: Comparison to Cas9 and NgAgo As you might expect given the...off-target editing and similar on-target editing when compared to the most commonly used Cas9 variant, SpCas9...Generating Mouse Models Using CRISPR/Cas9 Cpf1 Update: Comparison to Cas9 and NgAgo Special Delivery: Fluorophore...

  40. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...are of interest to you, our readers. Below I’ve compiled a list of the Top 10 Most Viewed Posts on our ...learn more about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating ...Morgan Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids 101: The Promoter...

  41. What's New in CRISPR - Winter 2018

    Type
    Blog Post
    ...difficult to compare the performance of different nucleases. Nicola Patron’s lab compared the efficiency...more predictable and precise deletion products compared to monomeric Cas9’s. Read the paper in Nature...

  42. 2022 and 2023: Reflections and Looking Forward

    Type
    Blog Post
    ...’d like to take a moment to reflect on our accomplishments from 2022 and share what we’re looking forward...ready-to-use recombinant antibodies collection, currently composed of over 100 antibodies. Everything in this collection...upon, a wealth of history to learn from, and a compelling future to plan in the changing landscape of science...

  43. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    ...tags with defined 3D structures less likely to compromise protein folding and function. Using bioinformatic...GFP did not reduce yeast cell size or viability compared to untagged GFP overexpression, while the rest...cell size and growth rate, indicating that they compromised protein-protein interactions. If you’re looking...

  44. A Conference By Postdocs For Postdocs: Future of Research

    Type
    Blog Post
    ...research and training is getting tighter and competition is fierce. However, I believe there are plenty...Washington University founded a successful consulting company called The Balsa Group. University of Pennsylvania... Consulting. I know of many schools that have completely student/postdoc-driven biotechnology clubs such...

  45. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...cassettes for selection of transgenic plants. The complete GreenGate toolkit, now available at Addgene, also...GreenGate can do is “save you half a year,” in comparison to more tedious and time-consuming traditional...traditional cloning methods, Lohmann said. The key component of Golden Gate and GreenGate are type IIS restriction...

  46. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ... For more details, check out this comprehensive review comparing optogenetic CRISPR techniques (Matony...repeatability. The system developed by the Sata lab is composed of two fusion proteins: 1) the genomic anchor ...of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies Publication...

  47. Bringing Sustainable Practices to the Lab: Innovations

    Type
    Blog Post
    ...labels are obtained by independent audit, and companies need to pay to obtain them. There’s a database...you don’t see one, be an ACTivist and encourage companies to obtain these labels! Reusing plastic Historically...benchtop, high throughput washing devices that are compatible with a variety of pipette tips. This is part ...

  48. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    ...bird feces, which get eaten by another snail, completing its lifecycle. Halloween genes The Halloween ...grasshoppers expressed different proteins in their brains compared to uninfected grasshoppers, including those linked...Additional resources on the Addgene blog Read about company culture at Addgene See some of our previous Halloween...

  49. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ...could take all of the credit, the success of the company is attributed to the entire Addgene family; the... on sliders, hot dogs and a mashed potato bar (complete with all the fixings), as well as plenty of delicious...course of scientific research. What was once a complicated process of requesting and sharing plasmids has...

  50. What's New in CRISPR - March 2020

    Type
    Blog Post
    ... Blue light renders the NLS accessible and the complex enters the nucleus where dCas9-Mxi1 can repress.... In the dark, the NLS is inaccessible and the complex remains in the cytoplasm. Read the paper in Cellular...CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout et al.,...
Showing: 501 - 550 of 929 results