Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

Information

In Category

We narrowed to 866 results for: cel.2

Showing: 361 - 380 of 866 results

  1. Deep Dive: qPCR

    Type
    Blog Post
    ...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor... that your reference genes are suitable for your cell lines and/or tissue types.  Please do note my oh-so-subtle...reference genes to save on plate space. For many cell lines and tissue types, appropriate reference genes...

  2. Spurred by COVID-19, Addgene Bridges Plasmid Sharing Between Academia and Industry

    Type
    Blog Post
    ...untagged expression of SARS-CoV-2 proteins in bacteria, yeast, and mammalian cells. We teamed up with Ginkgo...approximately 3,000 constructs to support SARS-CoV-2 research.  Browse the entire Ginkgo Bioworks COVID...organizations to share materials more broadly to accelerate COVID-19 research: Sinai Health System                                     ...

  3. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...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...

  4. Visualizing Translation at the Single Molecule Level

    Type
    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...

  5. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    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...

  6. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...you look at the dot plot in Figure 2, you might assume that all cell populations are positive for both ... the case of Figure 2, one population is B- and another two are B+. Our A-B+ cells are in the lower right...right corner.      Figure 2: This flow plot has two populations that are B+ and one population...-B+ cells, and A+B+ cells, but no cells that are A+B-.      Figure 3: It's often easier to identify...population of B cells and dendritic cells (DCs) in the lower middle right. DCs and B cells are HLA-DR+ and...single cells in a population. It can be used to measure protein expression, identify rare cells, or even...looking for cells in a sample that are negative for marker A and positive for marker B, or A-B+ cells. If you...

  7. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...starting day 2 after transfection.     Fig: 2: Expressing mARGAna in MDA-MB-231 cells. (a) Plasmid...expression. When engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...white inclusions in cells with GVs via phase contrast microscopy, GVs visible in cells via TEM, and purified... heterogeneity required single cell screening to isolate a clonal cell line capable of producing ultrasound...expression homogeneity, as the cells could be batch-sorted to isolate a polyclonal cell line with strong gas vesicle...transient transfection in rtTA+ cell lines (such as HEK293-TETOn cells), or they can also be integrated...

  8. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker pLenti CMV/TO GFP-Zeo...and display of proteins on the extracellular surface of S. cerevisiae cells His-tagged versions of pFastBac... the elements needed for replication in the host cell and are designed for a user to insert a gene of ..., or express the insert they carry in the target cell. These vectors allow you to test the function of....1-mCherry - Expresses mCherry tag in mammalian cells pBI-MCS-EGFP - Tet-inducible Find more Tet-inducible...or Fusion Protein Tags and fusion proteins are excellent tools for further understanding the function... as GFP, which allows you to visualize the cellular localization of your protein. Additionally...

  9. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...daughter cell, instead of partitioning each type of plasmid into a separate daughter cells (Figure 2) (Ebersbach...to plasmid loss (Diaz et al., 2015).  Figure 2: The partitioning problem. For low copy plasmids...Plasmids. Microbiology and Molecular Biology Reviews 62(2): 434–464. Pubmed PMID: 9618448 Diaz R, Rech J, Bouet...copy number in the cell is high, but allows replication when copy number in the cell falls too low (Novcik...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...eukaryotic cells). The partition NTPase is recruited and “walks” each plasmid to a separate daughter cell. When...from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...

  10. Hot Plasmids: Fall 2024

    Type
    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...

  11. Selecting Your Plasmid Purification Kit

    Type
    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...

  12. Droplet Digital PCR for AAV Quantitation

    Type
    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...

  13. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...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...

  14. Antibodies 101: Introduction to Immunofluorescence

    Type
    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...

  15. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...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...

  16. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...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...

  17. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser...ability to establish and maintain itself within host cells.  The plasmid has multiple ori that are functionally...eutrophus, Bartonella bacilliformis, Bordetella spp., Brucella spp., Caulobacter crescentus, Escherichia coli...

  18. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup...scFvs and T cell receptors and are expressed on T cells to make CAR T cells. The extracellular scFv portion...Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific antigen, it brings T cells to a ...recognizes an antigen of interest while the intracellular T cell receptor portion of the CAR facilitates ...scFv part of the receptor can activate T cells to kill cancer cells. scFv’s small size also allows for their...

  19. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...Zooming Forward Design: We produced a video celebrating Massachusetts Nonprofit Day, which was the perfect opportunity...  Quintin Marcelino...

  20. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...immunosurveillance." Cancer cell 30.1 (2016): 147-160. PubMed PMID: 27411589. 2. Todoric, Jelena, Laura Antonucci...represent a favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive...cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer, Inflammation and Immunity and...recent findings in basic immunology, cancer-immune cell interactions, cancer immunotherapy, as well as new...immunosurveillance. The presented study analysed a model of hepatocellular carcinoma, a cancer which can develop from ...example the upregulation of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment...follow the interactions between tumor and immune cells in real time. Addgene depositor Professor Max Krummel...
Showing: 361 - 380 of 866 results