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  1. To Each HIS Own

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    Blog Post
    ...will focus on one of the most common, purification based on the polyhistidine tag. What is the polyhistidine...tag was developed as a means to purify a protein based on the affinity of the imidazole ring of histidine...Bornhorst 2000).  One downside of polyhistidine tag-based purification, however, is the propensity for nonspecific... tags can also be used as a target for antibody based assays. Antibodies are useful tools for studying...
  2. Fluorescent Proteins 101: Fluorescent Biosensors

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    Blog Post
    ...Reporter signals may be electrochemical or light-based, with luminescent and fluorescent reporters being... domain induces changes in a fluorescent signal based on ligand presence or changing cellular conditions...Method Notes Quantification of a process Plate-based assays (plate reader) Detect the average amount...
  3. Twenty Years of Sharing Science!

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    Blog Post
    ...repository for DNA-based materials, including our popular kits and libraries. This DNA-based materials repository...
  4. TREAT-AD Plasmid Collection

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    Collection
    ...currently available. You can search or sort this table based on plasmid name, gene/insert, and more. ID Plasmid...
  5. TALEN Guide

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    Collection
    ... Figure 2 ). The Bogdanove group also hosts web-based software to help scientists find potential TAL effector...assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL...
  6. CRISPR in the Clinic

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    Blog Post
    ...initiating a base editor-based clinical trial for SCD in 2022 and Editas Medicine starting a Cas12-based clinical... today. Below we will review some of the CRISPR-based editing therapies that are currently approved or...
  7. 27 Hot Plasmids from 2016

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    Blog Post
    ...called GMAP (Gibson assembly-based modular assembly platform). GMAP is based on the Gibson Assembly method... Cas9-based AID tagging Seven years ago, Masato Kanemaki's lab successfully used the plant-based auxin-inducible...University, we are now providing a powerful CRISPR-based screening source for the Drosophila community as... it is composed of two sub-pools. The first is a base “90k library” that contains ~90,000 gRNAs that target... biosensor A pH-sensitive fluorescent biosensor-based assay system was recently deposited with Addgene...University of Zagreb, constructed a CRISPR-Cas9-based tool for targeting CpG methylation by fusing catalytically...proteins in a target cell. In addition to nucleotide-based linking strategies, post translational protein fusion...
  8. FastCloning: A Newer, Simpler Cloning Technique

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    Blog Post
    ...polymerase, the construction of specific sequences for base pair overhangs, insert and vector purification, ...specific to the insert with an additional 15-17 bases that are complementary to the overlapping vector...introduced into my inserts. However, with any PCR-based cloning technique there is a chance of introducing...
  9. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

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    Blog Post
    ...robust protocol it does have limitations. The enzyme-based CUT&RUN method overcomes many of these limitations...interacting. The CUT&RUN method Enter CUT&RUN, an enzyme-based protocol for identifying DNA binding patterns that...sets, of which there are many readily available. Based on the success that our lab and others have had ...
  10. Sensing Neuronal Dopamine

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    Blog Post
    ...from some labs have developed fluorescent-protein based DA sensors (Figure 2). The dLight1 dopamine sensor... dLight1 sensor is a single fluorescent protein-based sensor. This sensor was engineered from a GFP variant...sensor Yulong Li’s lab has created the GRABDA sensor based on a similar strategy: a GFP variant bound to a ...
  11. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

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    Blog Post
    ... or other products in the suite. Since it’s web-based, you could access it from anywhere. Its ease of ...Biorender is like clip art for scientists. The web-based collection of over 20,000 icons is designed by professional...stop struggling to make figures in PPT 🔬 Also, the base version is free, forever 💜 https://t.co/KJpyxgxeQt...
  12. Single-cell tracking of lineage and identity with CellTag

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    Blog Post
    ... level with CellTag CellTagging is a lentiviral based tagging method that labels cells with heritable ...three-fold increase of fully reprogrammed cells based on single-cell gene expression patterns. The lab...Chuner, et al. "CellTag Indexing: genetic barcode-based sample multiplexing for single-cell genomics." bioRxiv...
  13. Meet the New Addgene Board Members!

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    Blog Post
    ...School, he developed the first lentiviral microRNA-based system for polymerase II-regulated RNA interference...virology,” he says. He was working in an HIV lab and based on that experience, soon directed himself to finding...months, the lab began a project to develop an AAV-based COVID-19 vaccine.  Vandenberghe’s work with AAVs...
  14. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

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    Blog Post
    ...desktop-based software for plasmid design and mapping. ApE (A plasmid Editor): A free, donation-based plasmid...knowledge resource for cell lines IBEX Knowledge Base: A data repository for validation data on IBEX reagents...antibodies: A database of validated antibodies for mouse, human, and marmoset brain IHC.  FPbase: A data repository... Flame plasmid pET21a-BirA.   DNA / protein database search and analysis tools Addgene's analyze sequence...sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled from publications...nucleotide or protein sequences to sequences in a database ExPASy translation tool: Translates a DNA sequence...sequence using all six open reading frames UniProt: A database of protein sequence and functional information...
  15. 8 Great Depositor Research Stories

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    Blog Post
    ...tells us about a new tool for efficient multiplex base editing and multiplex prime editing (requiring minimal...dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation. AAV Q&A with Tim Miles...
  16. Adenoviral Vector Production and Troubleshooting

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    Blog Post
    ...therapeutic potential. Note: constructs discussed are based on Adenovirus type 5. Three generations of adenoviral...delivery: Potential applications for gene and cell-based therapies in the new era of personalized medicine...formation assay on A549 cells . Alternatively, antibody-based assays are commercially available for faster detection...
  17. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...short ncRNAs, such as microRNAs that are around 20 base pairs long. Using a single guide RNA (sgRNA) to ...Figure 1: A schematic illustration of CRISPR/Cas9-based approaches to study ncRNAs. (A) Transcription start...your experiments, there are plenty of other CRISPR-based options that can target ncRNAs.  CRISPRi and CRISPRa...10.1126/science.1225829  Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization...
  18. Uncovering Ancestral Bias in CRISPR Research

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    Blog Post
    ...American, Africa, or South Asian) used in the screens based on the cells’ germline genetic variations. They ...generating genetic dependency data with an ancestry-based bias.   The correction process does not stop there...genome-wide screens, introducing bias into all work based on these and any other CRISPR screens using the ...variation contributes to false negatives in CRISPR-based experiments with varying burden across ancestries...Cancer Dependency Map (DepMap), a cancer dependency database used to make this information accessible to the...
  19. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...packaging systems have been developed over the years based on HIV-1; however, the 1st-generation vectors are...defines 2nd- or 3rd-generation transfer vectors based on whether or not they have a chimeric 5'LTR. Second-generation...? What percent of the vector components are HIV-based? A7: No single plasmid contains all the components...insertional mutagenesis. These risks are largely based upon the vector system used and the transgene insert...
Showing: 181 - 200 of 706 results