We narrowed to 986 results for: IGH@;
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TypeBlog 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...
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Viral Vectors 101: An Introduction to AAV
TypeBlog 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... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog 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... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog Post...Lambert-Eaton myasthenic syndrome (LEMS) because, at high concentrations (25 mM), Gdm+ binds to and inhibits...disease-linked Arg-mutant proteins in the lab, clinicians might consider prescribing GdnHCl to patients suffering... -
Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog Post...and BioID, any protein that’s within a ~10nm neighborhood of the bait protein will be biotinylated. This...Macneill, S., & Varnaitė, R. (2016). Meet the neighbors: Mapping local protein interactomes by proximity‐dependent... -
Plasmids 101: Modular Cloning
TypeBlog 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... -
Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)
TypeBlog 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... -
Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog 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... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog 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 ... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog 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... -
Illuminating Epigenetics with A FRET Based Biosensor
TypeBlog 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... -
Gaining Leadership Skills Volunteering at a Professional Organization
TypeBlog Post...gradually. I didn’t become Chair of a Committee overnight (however this can be done if you already have ...was involved with postdoctoral issues first at Brigham and Women’s Hospital/Harvard Medical School and... -
How-to: 5 Steps to a Great Panel Discussion
TypeBlog 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... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post...gene knock-out (left) and targeted DNA knock-in (right) by CRISPR/Cas9. Adenoviral CRISPR vectors (left...left) or CRISPR vectors and donor DNA vectors (right) are transduced to target cells. Within target cells... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog Post...cluster of enhancers. CAPTURE 2.0’s multiplexing and high resolution allowed the Xu Lab to simultaneously ... chromatin's dynamic role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog 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... -
Isolating Genomic Regions of Interest with the CRISPR System
TypeBlog Post...purified by streptavidin high-affinity purification. This system is both highly sensitive and specific ... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog Post...current period of rapid flux, the chapter might still feel slightly dated by next week. A stepping stone... -
Viral Vectors 101: The Retroviral Lifecycle
TypeBlog 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... -
A Guide to Starting Your Own Journal Club
TypeBlog 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 ... -
Simplify Cloning with in vivo Assembly
TypeBlog 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... -
Lentiviral Vector Uses and Overview
TypeBlog 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... -
A Career in Grants: How To Become a Grant Professional
TypeBlog 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... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog Post...around 2010 and found the scheme very nice at first sight in that Addgene takes care of everything including...research to realize their vision. Their progress might be slow, but if they don't stop, they may be able... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog 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... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...following injection into the bloodstream, but requires a high vector dose and is unable to reach all regions of...small genetic difference between these two mice might drive their divergent phenotypes. Two labs, the ... -
CRISPR-mediated Plant Base Editors
TypeBlog 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... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog Post...Syndrome research, the role of the MECP2 gene, and highlights some of the latest breakthrough therapeutic approaches... -
To Codon Optimize or Not: That is the Question
TypeBlog 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... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog 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... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog Post...positive cells isolated with TREE have a significantly higher frequency of genomic base pair edits of interest...positive cells can be sorted and have a significantly higher frequency of base edits of interest than solely... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog 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... -
Bioinformatics at Addgene
TypeBlog 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... -
Plasmids 101: Shuttle Vectors
TypeBlog 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... -
CRISPR 101: Any Base Transversion Editors
TypeBlog 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... -
Bacterial Toxin-antitoxin Systems as Molecular Biology Tools
TypeBlog Post..., cells that retain the ccdB gene die off and a higher percentage of transformants contain the ccdB-less...great additions to constructs that are used to drive high protein expression in vivo. Expression plasmids ... -
Plasmids 101: Control Plasmids
TypeBlog 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... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog Post...plasmids expressing variants with optimal function. High-throughput DNA sequencing is then used to measure...mutant strand: A mix of degenerate primers and the high-fidelity Phusion polymerase are used to introduce... -
CRISPR 101: Homology Directed Repair
TypeBlog 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... -
Using Phosphoserine to Study Protein Phosphorylation
TypeBlog 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...