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We narrowed to 595 results for: cel.2;

Showing: 261 - 270 of 595 results
  1. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ... than control, 3; same as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice...expanded since its adaptation for use in mammalian cells in 2012-2013. Researchers are now using this system...plasmids containing sgRNA sequences into HEK293T cells. The fluorescence intensity was classified into ...is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with...incubation, you assess the level of fluorescence in your cells under a fluorescence microscope. The higher the ...
  2. Selecting Your Plasmid Purification Kit

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    Blog Post
    ... environmental sustainability in mind (Table 2). Table 2: Sustainability features highlighted in plasmid...plasmid DNA from unwanted cellular “debris”; and then clean-up the plasmid DNA (Figure 2). For most kits, the...together — no mixing and matching!   Figure 2: Plasmid isolation workflows are similar across kits...the steps for lysing cells and removing cellular debris look very similar. However, keep in mind that ...relative term — even armed with the highest-competency cells you will still struggle to get transformants if ...quantity) – Depending on the experiment, mammalian cell transfections can require quite large amounts of...your bacteria to release the plasmid, separate the cell debris from the plasmid, and then clean-up the plasmid...
  3. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

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    ...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...Xenopus frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then ...is the smallest photosynthetic cell in existence, but most abundant cell on earth (twice the human biomass...only 1,700 genes total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little...Mormyridae (Jason Gallant) Weakly electric fish are excellent systems to study for convergent evolution because...
  4. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...into the several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, ...mounting sample on slides, and confocal   Figure 2: High-level overview of an indirect immunofluorescence...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment...interest live within a cell,” “Does this disease change the architecture of my cells,” or “How does this ...IF), is an immunoassay that brings to light the cellular world. The technique allows you to ask questions...this mutation impact the types of cells found in my tissue.” It is based on the same principles as other ... visualize a target of interest within an intact cell, tissue, or, in some cases, a whole organism. But...
  5. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    ...Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Find...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...are capable of cleaving genomic DNA in mammalian cells and zebrafish embryos, and they can be used to modify... adapted for targeted RNA cleavage in mammalian cells. The REPAIR (RNA Editing for Programmable A to I...reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol 34:863–868 . https://doi.org/10.1038...specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol 34:869–874 . https://doi.org/10.1038...
  6. Visualizing Translation at the Single Molecule Level

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    Blog Post
    ...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor... of all of the mRNA bound-ribosomes in a cell. When cellular mRNAs are digested with ribonucleases, ribosomes...Regulating translation is key to cellular function, especially during development or stress. With ribosome...explore the complexity of translational control in cellular physiology. Ribosome profiling Ribosome profiling...the effects of 5’ UTR variants in translation of cell cycle protein Emi1, showing that ~80% of transcripts...powerful tools for studying translation in various cell types and physiological states. References 1. Ingolia...
  7. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

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    Blog Post
    ...Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence...plant protein TIR1 in non-plant cells, which interacts with the cells’ endogenous ubiquitin ligase, the...translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al., 2016...based method to tag endogenous proteins in HCT116 cells with mAID. Using this method, the lab was able to... protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that...the target sites adjacent to an NG PAM in human cells. The increased versatility of the engineered SpCas9...
  8. Hot Plasmids: Fall 2024

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    Blog Post
    ...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...useful addition to the toolkit for visualizing cellular processes in C. elegans: mScarlet3 for applications...biologically relevant forces at focal adhesions in living cells in several different contexts.   Figure ...substrate furimazine. Right: Demonstration of PILATeS. Cell adhesion to a PILATeS-functionalized coverslip creates...
  9. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...amplitude measured for each droplet in each well (Figure 2). A clean ddPCR should have a clear separation between...as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has ...used in the detection of rare sequences and single cell analysis. It is also being used in the detection...
  10. Antibodies 101: Introduction to Gating in Flow Cytometry

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    ...represents the time and intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength...most people use that setting. Figure 2: As the cell passes through the laser beam and causes it...proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line one ...Many cell types can be found in the tumor microenvironment (tumor cells, immune cells, blood cells and ...about 53% of my cells are dendritic cells and ~13% are CD8+ T cells. Around 34% of the cells are neither ...neither of the two cell types (likely B cells and others). Almost no cells stain positively for both markers...measure your cells of interest. In many experiments you’ll be working with a heterogeneous cell population...
Showing: 261 - 270 of 595 results