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  1. Addgene: The First Twenty Years

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    Blog Post
    ...viral vectors were useful for researchers wanting high quality reagents, and we realized we could use the...Addgene’s support, could fill a community need for high-quality service, open data, and increased reproducibility...
  2. Viral Vectors 101: An Introduction to AAV

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    Blog Post
    ....7 kb. Co-infection rates for AAVs are generally high enough for this approach to work, although it’s ...time. Though the time delay until expression is highly dependent on the serotype and target tissue, two...
  3. Fluorescent Proteins 101: Introduction to FRET

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    Blog Post
    ...1948 as a variation of the more commonly observed light emission by fluorescence. The widespread use of ...distances that are too miniscule for conventional light microscopy. Don't FRET - Use these FRET resources...
  4. Plasmids 101: Modular Cloning

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    Blog Post
    ...of large-scale assembly useful, you may ask? You might be surprised by how multi-purpose it can be. Let... Plant Parts Kit Resources on the Addgene blog High Complexity Golden Gate Assembly with NEB’s DAD Interview...
  5. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

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    Blog Post
    ... is biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated to the...interest. The optical density (OD) - the measure of light absorbed by a solution - of these wells is plotted...
  6. Adeno Associated Virus (AAV) for Cell and Gene Therapy

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    Blog Post
    ...are a multitude of known AAV serotypes that have slight variations in their viral capsids. Novel forms ...AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the human population. These pre-existing...
  7. Adapting Toehold Switches to Detect Zika Virus

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    Blog Post
    ...for these RNA sequences (3 nM), however, was quite high and above what would be expected in patient samples...daunting. It took a true team effort and many late nights, not unlike most research, but the urgent need ...
  8. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and should be stored...resistance marker expression following transfection, higher-level expression after approximately 72 hours is...
  9. Illuminating Epigenetics with A FRET Based Biosensor

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    Blog Post
    ...expression can change as well. The more frequently and tightly bound DNA is to histones, the more compacted it...occurs at CpG islands (regions of the DNA with a high frequency of C and G nucleotides). The conversion...
  10. How-to: 5 Steps to a Great Panel Discussion

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    Blog Post
    ...my experience the best set-up is panelists in a slight semi-circle (so they can see each other) on comfortable...redirect the conversation. The person you cut off might be miffed but the other 200 audience members will...
  11. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...think I need. Unlike lentivirus, AAV requires a higher number of viral particles for efficient gene transfer...only harvest the cell pellet. My yield would be higher if I used PEG precipitation, but for my experiments...
  12. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ...Gamma-retrovirus genome (left) and an HIV lentiviral genome (right.)  HIV seems an unlikely candidate for use as...genomes in one retroviral particle: pseudodiploidy and high rate of genetic recombination. Proceedings of the...
  13. A Guide to Starting Your Own Journal Club

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    Blog Post
    ...wanted to think more closely about the wealth of high-throughput sequencing (HTS) data we generate and...the aim of the journal club is to begin building a high level understanding across multiple teams or to ...
  14. Simplify Cloning with in vivo Assembly

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    Blog Post
    ...bp, but pay attention to the sequence’s Tm, as a higher Tm give more efficient recombination. We normally... two colonies for simple cloning procedures. Use high fidelity DNA polymerase This will minimize random...
  15. Lentiviral Vector Uses and Overview

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    Blog Post
    ...cumbersome to use and lead to lower viral titers due to higher plasmid number. For more information on the key...Lentiviral particles with the VSV-G envelope have high infectivity in a wide range of cell types, so they...
  16. A Career in Grants: How To Become a Grant Professional

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    Blog Post
    ...professionals identify grant opportunities with the highest likelihood of funding and develop a strategy and... project management, and program evaluation are highly transferable. Use your experience and your passion...
  17. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...to Cas9, which requires that each DNA target have high sequence identity to the spacer sequence and contain...to detect single molecules of an RNA species with high specificity (Gootenberg et al., 2017). This system...
  18. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...This post was contributed by Kutubuddin Molla, a Fulbright Visiting Scholar at the Pennsylvania State University...Cuttack, Odisha, India. Currently Kutubuddin is a Fulbright Visiting Scholar at the Pennsylvania State University...
  19. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...? Let’s take a look at codon usage and when you might want to optimize codons for expression in a particular... on the codon usage frequency in a reference of highly expressed genes (Sharp and Li, 1987). In 2000 the...
  20. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ... method, they found that SaCas9 did not display higher levels of off-target activity than SpCas9, confirming...editors. While using AAV to deliver large transgenes might seem daunting, it’s actually a challenge that the...
  21. Twenty Years of Sharing: Expanding Accessibility

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    Blog Post
    ...which help a researcher understand if a material is right for their experiment.     Table 1: Most popular ...learned to clone from the Addgene website.” We delight in seeing our resources shared via social media...
  22. Bioinformatics at Addgene

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    Blog Post
    ... was to ensure the scientific community obtained high-quality data for our DNA-based reagents. Not only..., one of our first problems was deciding on the right system for us to start using. Prior experimenting...
  23. Plasmids 101: Shuttle Vectors

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    Blog Post
    ...eukaryotic ribosome. One resistance gene on your vector might require two different selection molecules! It all...protein, to visualize vector uptake. These reporters might be used as markers for automated cell sorting or...
  24. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ...Although AYBE, AXBE, and ACBE do not approach the high product purity of CBEs and ABEs, they could be useful...transversion editors, this editor had relatively high indel rates ranging from 4.7% to 30% at tested sites...
  25. Plasmids 101: Control Plasmids

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    Blog Post
    ... you are trying to repress a gene, your control might be a system where expression of that gene is knocked...selection of the positive control plasmids should be highly specific to the experiment and the interrogation...
  26. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...ionizing radiation, chemotherapeutics, and ultraviolet light. In this post, we will describe the mechanism of...systems, ssODN templates generally provide the highest frequency of HDR edits.  Check out our blog post...
  27. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...,000 phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the...intersection of cell signaling, metabolism, and the mighty mitochondrion. You can follow her on Twitter @nieminm...
Showing: 801 - 840 of 986 results