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We narrowed to 313 results for: POR C

Showing: 261 - 280 of 313 results

  1. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...doi.org/10.1038/nprot.2007.267 Stoeckius M, Hafemeister C, Stephenson W, et al (2017) Simultaneous epitope and...find yourself doing your own conjugation, it is important to keep in mind that you will need to confirm ...

  2. Plasmids 101: Modular Cloning

    Type
    Blog Post
    Published
    March 12, 2024, 1:15 p.m.
    ...References and Resources References Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. A modular cloning...phenotype, translational output, etc., which can support a larger cellular engineering goal, several of ...

  3. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    Published
    Nov. 19, 2015, 3:30 p.m.
    ...islands (regions of the DNA with a high frequency of C and G nucleotides). The conversion of cytosine to ... versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27...The K9 reporter uses the HP1 chromodomain to recognise lysine 9 methylation, while the K27 reporter uses... methylation in living cells. Essentially this reporter construct consists of one substrate (the histone-derived...-K27 histone methylation reporter) positions on histone H3, which are known to be involved in repression...methylation of these sites was controlled, the reporter consistently showed significantly different FRET...states. Thus, this system is a reliable tool in reporting methylation status of these sites. This new technology...

  4. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ... PubMed Central PMCID: PMC3622210. Schek N, Cooke C, Alwine JC. Definition of the upstream efficiency ...expression levels, the terminator also plays an important role in RNA processing and contributes to variability...polyadenylation. The addition of the poly(A) tail is important for stability of the mRNA, protection from degradation...degradation, and is integral to the nuclear export and translation processes as well.   Figure 2... non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and/or use...the Mason–Pfizer monkey virus constitutive RNA transport element lead to enhanced HIV-1 Gag expression ...

  5. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ...procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can be used to confirm...Professor at Boston College. National reports stress the importance of providing authentic research experiences... is particularly interested in strategies to incorporate research into undergraduate courses and in communicating...

  6. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ... Cas9 to generate the mutant cells, respectively (C). Mutant cells are screened (D) and "hits" are identified...post was updated on Aug 20, 2020. What genes are important in your phenotype of interest? Many scientists...not entirely known. Identifying which genes are important for a phenotype can lead to a wealth of additional...strong evidence that a particular gene is actually important for a given phenotype. Each hit from the screen...

  7. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ...//doi.org/10.1016/s0091-679x(08)60255-1  Manning, C. F., Bundros, A. M., & Trimmer, J. S. (2012). Benefits... reactive ones. And make sure to check for any reported cross-reactivity when planning your experiment...Why Choosing the Right Secondary Is of Primary Importance. In J. Sturtevant (Ed.), PLoS ONE (Vol. 7, Issue...

  8. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which result in the internalization...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane...and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target effects can ...through a process known as CRISPR-EZ (CRISPR RNP Electroporation of Zygotes) (Chen et al. 2016). For CRISPR ...CRISPR experiments that involve HDR, electroporation can be combined with cell-type specific reagents in a ...a technique known as nucleofection, which forms pores in the nuclear membrane, allowing for entry of a... account with these types of experiments. One important consideration is that the Cas9-gRNA RNP will be...

  9. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...media, reducing the incubation temperature (e.g., 30 °C), and limiting your growth time (e.g., 16-18 hours...individual libraries can vary significantly, it’s important that we have an amplification protocol developed...and the desired coverage for the library (How important is it that every single plasmid in your library...protocol. Created with BioRender.com.   The most important factor to consider is your desired library coverage...not millions, of plasmids, they are generally electroporated into electrocompetent cells to ensure the highest...chemical transformations is less than that of electroporation, but an advantage is that chemical transformations...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...

  10. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist ...a fluorescent emission wavelength in the green portion of the visible spectrum (hence the name), which...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...solvent. This tightly-packed structure explains the importance of the entire GFP protein, which is almost completely...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest,...

  11. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ... targeting mCherry reporter (mChe), with co-incubation with assist peptide (AP). C) Workflow for Cas-RNP-PAGE...difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection.... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized by pre-existing antibodies...

  12. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha N, Gil S, Wang H, Papayannopoulou T, Lieber...pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR activity... way to make those genome edits. But, just as important is a way to flip the switch and turn off CRISPR...The ability to turn off CRISPR activity is an important step towards CRISPR-based therapies and for more...

  13. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...     References and Resources References Shen C-H (2019) Amplification of Nucleic Acids. In: Diagnostic... a fluorescent signal. The signal increases proportionally to the amount of product produced each cycle...multiple starting DNA templates in one sample. It is important to note however, that multiplex PCR is more complicated...

  14. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011...have that conversation out in the open is really important. Addgene: Is there anything else you’d like to...biology is massively underfunded considering how important it is. I would really like to reach all those ...

  15. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...4. Mittereder, Nanette, Keith L. March, and Bruce C. Trapnell. "Evaluation of the concentration and bioactivity... quickly due to immune response against AdV. Importantly, recombination-competent adenovirus (RCA) can...

  16. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab found a way around this by using ... Signaling and Protein Purification RANbodies: Reporter Nanobody Fusions Plasmids 101: Secondary Nanobody...

  17. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ... where they could bind to intracellular epitopes. C) Permeabilizing with a non-ionic detergent, like Triton... for antibodies that bind to the extracellular portion of a protein. B) Permeabilizing with the selective...

  18. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ... living cells. Shao S, Zhang H, Zeng Y, Li Y, Sun C, Sun Y. Commun Biol. 2021 Yujie Sun   For even...of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin. Lu et al. Plant J. 2010... (abundant in eukaryotes) A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein Interactions...green-yellow or orange-red fluorogen A split fluorescent reporter with rapid and reversible complementation. Tebo....  Controls  When designing experiments, it’s important to include proper controls to assess the behavior...target proteins, especially if the termini are important for their interaction. You can read our previous...

  19. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia R, Marillonnet S. A one pot, one step, precision...produce complex fluorescent reporter constructs. These fluorescent reporters can be used to detect structure...), the same cloning procedure can be used to incorporate each element into the final construct. This strategy...protein sequences to develop diverse fluorescent reporters is a useful application of high-throughput cloning...sequences), so enzyme choice is critical. May not support in-frame fusions of coding sequences Sladitschek...

  20. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...Moreno-Mateos, M. A., Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed...typically require an exact DNA sequence to be incorporated at a precise genomic location, with little to...template for repair, which will result in the incorporation of your sequence, a.k.a a knock-in! So how does...higher HR frequencies than quiescent cells. It is important to know that HR is often mutated in cancer cells...molecule is used as the template, you need to incorporate homologous sequence to the right and/or left ...
Showing: 261 - 280 of 313 results