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We narrowed to 718 results for: des.1

Showing: 441 - 460 of 718 results
  1. Plasmids 101: Protein tags

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    Blog Post
    ...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function... conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based ... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the...depending on their protein of interest. After several decades of trying to address these challenges, researchers...examples. The tables list more common tags than are described in the post, but have been categorized to help...
  2. To Codon Optimize or Not: That is the Question

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    Blog Post
    ... cerevisiae (Tuller et. al., 2010). Figure 1: An overview of peptide synthesis. A ribosome interacts...McClain WH (2000) The G·U wobble base pair. EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001...Usage Tabulated from GenBank database (CUTG), that provides an electronic dataset for codon-usage of 257,468...(A) The codon wheel depicts the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example...Codon Optimization  ATUM DNA2.0  IDT Synthetic gene design made easy GenScript- GenSmart Codon Optimization...
  3. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with...follow-up posts: (1) (2). Base editors have a narrow editing window of only a few nucleotides, so they are ...or one pyrimidine base to the other. Figure 1: Base transition editors catalyze purine-to-purine...engineering and protein delivery. Nature Communications, 8(1), 15790. https://doi.org/10.1038/ncomms15790 Richter...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...
  4. Antibodies 101: The Four ELISAs and When to Use Them

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    Blog Post
    ...alternative detection methods.      Figure 1: The four types of ELISA are direct, indirect, sandwich...conjugated antibody is not readily available. Table 1: Overview of the advantages and disadvantages of each...staining. Antigen-containing samples often include undesirable molecules that may bind non-specifically to assay...specific than other methods. In addition, the assay includes both primary and secondary incubation steps, increasing...
  5. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...Spaghetti Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins... expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes...map these neuronal networks. For almost three decades, recombinant Adeno-associated virus (AAV) has been...symbol of opposition to the teaching of intelligent design in public schools and was started in 2005 in Kansas...
  6. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    Blog Post
    ...cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can block CRISPR activity... ways to anti-CRISPR proteins in your experiments 1. Reduce off-target effects Prolonged Cas activity .../35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity in Human Hematopoietic...
  7. Antibodies 101: Flow Compensation

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    Blog Post
    ... present in the sample (APC in the case of Figure 1). In Figure 1A, the population needs to be shifted... decrease the compensation value.   Figure 1: Compensation example with single color control. Flow...This will make it difficult to properly gate your desired cell population. By compensating between fluorescent...
  8. 10 Basic tips for mammalian cell culture

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    Blog Post
    ...for example, split cells on a Monday diluting them 1:10, you should be able to split on the following Thursday...Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis...and could be contaminated.Learn how Addgene uses barcodes to track samples Keep track of passage numbers...
  9. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene...the years. The authors optimized these systems by: 1) decreasing the number of plasmids required for each...
  10. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...by insertion and deletion frequency. Fig. 1: Overview of TIDE. The input is sequencing trace files...keep your product to knock-in size ratio below 10:1. Small knock-ins Is your knock-in edit small, with...screening by PCR may be the way to go. Design a PCR to flank your desired edit site and run the PCR product...nucleotide along with your desired edit. This silent mutation should be designed solely for restriction enzyme... updated Nov 3, 2022 by Susanna Stroik. You’ve designed your gRNA and introduced it into your target cells... your cells were appropriately edited for your desired mutation - insertion, deletion, or site-directed...know a few things about your target cells and the desired edits. First, you will need to know what the ploidy...
  11. Site Directed Mutagenesis by PCR

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    Blog Post
    ...mutation (usually 1-3 mismatched bases) is sufficient for your primers to successfully anneal to the plasmid...researcher at Cornell University.       References 1. Breslauer, Kenneth J., et al. "Predicting DNA duplex...for the desired modification. Finally, the positive clones are sequenced to confirm the desired modification...Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the desired mutation...introduced to plasmids using primers (with the desired mutation) in a PCR protocol that amplifies the ...mutated clones. Forward and reverse primers are designed to be complementary, but each primer may extend...Polymerase To ensure that you don’t introduce undesired mutations through the PCR process, you need a ...
  12. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ...folding, and deal with sub-optimal codon usage (Table 1) A few mutations are common to all or most expression...curated list of Bacterial Expression Systems Table 1: E. coli expression strains  Note: All strains are...and some of the most popular commercial strains designed to avoid them. Why do I need an expression strain...protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce proteolysis of the...which can prevent the culture from reaching the desired density within a reasonable time-frame. For these...and pLysE—the difference being that the latter provides tighter control of basal expression. What if I...don't see protein overexpression? The strains described above should generate sufficient expression levels...
  13. Streaking for Single Colonies: The Streak Plate Challenge

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    ...Time for the #BioSci3319 #StreakPlateChallenge part 1. Lab B needs a redo :-( @SandTBioSci #CompetitiveStreaking...the Streak Plate Challenge, Westenberg’s course includes Winogradsky columns and characterizing the microbes...
  14. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

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    ...know that most cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult...organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...promotes scientific sharing The EDGE Program is designed to expand the number of systems that can be studied...genotype to phenotype studies New organismal systems described at this meeting were extraordinarily diverse. ...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...control who a female wants to mate with and if she decides to raise their offspring or leave them to their...
  15. New Viral Vectors - Spring 2025

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    Blog Post
    ... new preps might enhance your research!   Table 1: New ready-to-use viral vectors in the Addgene repository...community and support a wide range of experimental designs. Here are nine great additions, including our first...
  16. The Addgene Antibody Data Hub

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    ...an antibody performed in an experiment.   Figure 1: The front page of the Addgene Data Hub.   Antibody...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...
  17. CRISPR Activation: A Practical Guide

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    ...activity to a transcriptional activation domain (Fig 1). Two ways to use CRISPRa for your research CRISPRa... Stucka R, Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional...whichever guides are closest to the transcription start site (TSS). We recommend targeting the guides to a ... you should also be ready to test three or four guides directed towards that gene as there can be a large...transcription. That being said, we generally pick guides by putting the promoter of the gene into a gRNA...
  18. Addgene's Website Update

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    Blog Post
    ...straight to your materials of interest!   Figure 1: The new search bar.   Pro tip! You can use a manuscript...and the Addgene Help Center. A big part of this redesign was pulling all of our science options into a ...Addgene’s website easier to navigate, thanks to design choices informed by Web Content Accessibility Guidelines...word for it — check out all the new features and designs addgene.org! And when you’re finished, feel free...
  19. Your Top Requested Plasmid in 2015!

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    Blog Post
    ...forward to another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...Feng Zhang’s lab at the Broad Institute and was designed and cloned by Neville Sanjana. This fantastic ...your gRNA of interest to mammalian cells. It was designed to have increased lentiviral titer as compared...sampling of the research it has already been used for includes work probing mTOR signaling in cancer cells, deciphering...
Showing: 441 - 460 of 718 results