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We narrowed to 1,035 results for: URE

Showing: 101 - 120 of 1035 results

  1. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ... manipulate biology based on molecular features found in nature, with the ultimate goal to improve the...impact on this challenge either for magnetic capture or for capture using microfluidic devices. Addgene: What...antibodies against common tumor markers to subsequently capture cancerous cells expressing low levels of tumor ...binding stability really mattered for efficient capture. We wanted to be able to build a highly multivalent...for keeping going while we worked out the cell capture system.) Cancer and other applications Addgene:...technology mean in the context of cancer? Howarth: The capture of circulating tumor cells (CTCs) is going to have...treatment of cancer. It is clear that one needs to capture a diverse range of CTCs to cope with the variation...

  2. CRISPR Guide

    Type
    Guide
    ...addition of a structured motif to enhance pegRNA stability PEmax - optimized PE architecture; increased ...synthesized (Figure 8A), then cloned in a pooled format into lentiviral transfer vectors (Figure 8B). Libraries...Berkeley. Plan Your CRISPR Experiment Resources Figures All figures were created using BioRender.com (Link opens..., N., Ishitani, R., Zhang, F., & Nureki, O. (2014). Crystal Structure of Cas9 in Complex with Guide RNA...vivo genome editing using Staphylococcus aureus Cas9. Nature , 520 (7546), 186–191. PMID: 25830891 Rauch...became the most popular genome engineering approach. Figure 1: Overview of the basic CRISPR mechanism Engineered...gRNA left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will...

  3. Antibody Guide

    Type
    Guide
    ...isolated and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is...Antibodies can also be produced in cell cultures using hybridomas, cell cultures created by fusing B cells with...form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for binding the...Monoclonal antibodies are generated from cultured B cells, where each culture should produce the same antibody...Diabodies can be used for assembling protein nanostructures. Figure 2: An antibody (IgG, far left) compared... a signaling molecule. Figure 4: A direct sandwich ELISA, which uses a capture antibody and a primary ...In a sandwich ELISA, a capture antibody, bound to a surface, is used to capture the protein of interest...

  4. Lentiviral Vector Guide

    Type
    Guide
    ...Table 1. Figure 4: Third-generation lentiviral plasmids. Created with BioRender.com. Feature Second-generation... chromatin architecture through large-scale mapping of lentiviral integration sites. Nature Protocols ...to continue their lifecycle and replicate. This feature can be harnessed for stable delivery of various... typically ranges from 8–10 kb, encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus...have been removed or mutated for increased safety. Figure 1: Wild-type lentivirus genome. Created with BioRender.com...largely intact. First-generation plasmids include (Figure 2): Transfer plasmid — contains transgene and wild-type...envelope removed Envelope plasmid — contains env Figure 2: First-generation lentiviral plasmids. Created...

  5. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...replication and also act as signals for packaging. Figure 1: Wild-type AAV genome. Created with BioRender.com...do not trigger a strong immune response. These features have made AAVs the preferred choice to be used...components, see the AAV Plasmid Elements table . Figure 2: Plasmids required for rAAV vector production...methods (such as a Gateway destination vector), so be sure to confirm your chosen plasmid is compatible with...available in your lab. When cloning your plasmids, be sure to use recombination deficient bacterial strains... the ITRs found in transfer plasmids. This will ensure that the repeats will be maintained and often results...and instead provided by the chosen cell line to ensure safety and prevent the potential formation of replication-competent...

  6. Plan Your Experiment

    Type
    Guide
    ...different optimal editing windows. Be sure to refer to the literature associated with your chosen editor....activity. Be sure to sequence the genomic region you intend to target in your samples to ensure there are...components (Figure 2), and the optimal system will likely depend on your cell type. Figure 2: Different...(Cas enzyme and guide RNA) for your experiment (Figure 1). You will decide how to express Cas9, the delivery... and finally how to validate your genetic edit. Figure 1: Flow chart describing the general framework ...and essentially function as a “knockout,” but be sure to consider potential consequences of having a non-functional...only compatible with smaller Cas enzymes, like S. aureus Cas9. For more information, see our blog post on...

  7. Adenovirus Guide

    Type
    Guide
    ...Collection page . Figure 5: Overview of AdEasy™ adenoviral vector production system. Figure is based on the...assemble icosahedral capsids and build new virions. Figure 1: Wild-type adenovirus and its genome. Genes over...which incorporate additional genomic deletions. Figure 2: First-generation rAdV vectors. Created with ...using first- and third-generation rAdV vectors. Figure 3: Second-generation rAdV vectors. Created with... challenge of the HCAdV vector production is to ensure no carryover from the HV in final viral preparations... genome or using other plasmids instead of HVs. Figure 4: Third-generation rAdV vectors and helper virus...methods (such as a Gateway destination vector), so be sure to confirm your chosen plasmid is compatible with...

  8. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...to continue their lifecycle and replicate. This feature can be harnessed for stable delivery of various...gamma-retroviruses ranges from 9–11 kb, encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus...producing viral vectors in a laboratory setting. Figure 1: Wild-type gamma-retrovirus genome. Created with...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA...infectivity for different cell types (known as tropism). Figure 2: Gamma-retroviral plasmids. Created with BioRender.com...methods (such as a Gateway destination vector), so be sure to confirm your chosen plasmid is compatible with...available in your lab. When cloning your plasmids, be sure to use recombination-deficient bacteria strains,...

  9. Optogenetics Guide

    Type
    Guide
    ...Guide to Channelrhodopsin Variants: Features, Limitations and Future Developments. Exp Physiol. 96(1): ...optogenetics integrates optics and genetic engineering to measure and manipulate cells (frequently neurons) and their...optogenetics research utilize light to detect, measure, and control molecular signals and cells in order...functions. Example schematic of an optogenetics procedure. A channelrhodopsin, fused to mCherry, is expressed... allow the fast depolarization of neurons upon exposure to light through direct stimulation of ion channels... in excitation (depolarization) of the neuron. Feature enhancements include: Increased photocurrent amplitude...from the cryptophyte Guillardia theta ). Other feature enhancements include: Increased photocurrent amplitude...

  10. Chemogenetics Guide

    Type
    Guide
    ...check out our Chemogenetics Plasmid Collection . Figure 1: General overview of a chemogenetics experiment...muscarinic receptor and has not been used in vivo . Figure 2: DREADDs, their ligands, and signaling properties...and/or K + ions, and activates neuronal activity. Figure 3: PSAM4-based ligand-gated ion channels, their...optogenetic systems in our Optogenetics Guide . Figure 4: Luminopsins can be activated through chemogenetic... control expression of a chemogenetic reporter ensures that the reporter remains silent until a cell or... X, Stone O, Roth BL, Jin J (2015). The First Structure–Activity Relationship Studies for Designer Receptors...

  11. Molecular Biology Reference

    Type
    Guide
    ...propagation of these plasmids through cell division. To ensure the retention of plasmid DNA in bacterial populations.... Under these conditions, there is a selective pressure to retain the plasmid DNA, despite the added replication...organisms, including plants, worms, mice, and even cultured human cells. Although plasmids were originally...bacteria, but also provide those bacteria with a pressure to maintain and replicate your plasmid over multiple... extremely important when verifying plasmids to ensure each plasmid contains the essential elements to...replicated using the modified bases and a microscope captures the fluorescent color that is emitted each time...

  12. Science Guides

    Type
    Guide
    ...conjugated to a signaling molecule, can be used to capture, quantify, and identify proteins of interest. Read...Optogenetics integrates optics and genetic engineering to measure and manipulate cells (frequently neurons) and their...optogenetics research utilize light to detect, measure, and control molecular signals and cells in order...

  13. Addgene's eBook Collection

    Type
    Guide
    ...Not sure where to start? Need a quick reference? Addgene's eBooks are here to help!... Collection Not sure where to start? Need a quick reference? Addgene's eBooks are here to help! 0 of 0...you need to know about plasmids, from parts, nomenclature, and uses to common promoters and cloning techniques...

  14. Guide to Using Pooled Libraries

    Type
    Guide
    ...infected cell receives only one plasmid. To make sure that every plasmid is adequately represented in ...planning to conduct a pooled library screen. The figure above is an outline of how a gRNA library is used...cells that survive post-selection. The selective pressure must be strong enough that most of the cells die...

  15. Modular Cloning Guide

    Type
    Guide
    ... a protein-coding gene, and a terminator, as in Figure 1). Next, up to six Level 1 modules can be assembled...maintain the final plasmid after transformation. Figure 1: Overview of the MoClo Toolkit components and...synthetic biology. MoClo Plant Parts II and Infrastructure Kit Plant Expression Johannes Stuttmann 95 ...

  16. Promoters

    Type
    Guide
    ...the TSS. Eukaryotic promoters are so complex in structure that the DNA tends to fold back on itself which...which is a main component of a cell’s ribosome structure. Ribosomes are the site of protein syntehsis where...

  17. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...crystallography cannot capture flexible or disordered regions, so crystal structures like those in Figures 2 and 3 don...biology! Major features First, we’ll cover the basic parts of the Cas9 enzyme.   Figure 1: A cartoon...from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state...state that it doesn’t appear in the crystal structure (Figure 2) at all. When this crucial recognition region...learn about this process!      Figure 3: Crystal structure of S. pyogenes Cas9 bound to guide RNA...are disordered and don’t appear in the Figure 3 crystal structure, reflecting the less stringent recognition...tolerate mismatches in this region. The crystal structure in Figure 3 shows the guide RNA stabilizing the DNA-binding...

  18. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...contamination of the rest. Avoid long exposure of the cell culture to air and make sure to tighten the flask cap before...contamination in your cell culture or you are about to begin a new cell culture project. If mycoplasmas ...They are present in most of the cell culture facilities, tissue culture labs and every cell culturist has... every cell culture. Reusing the same bottle of serum again and again for each subculture can enhance ...does not ensure complete sterilization as mycoplasma can even escape 0.2-micron filters. Figure 1: Contamination...seen in the culture, it is a good option to discard the flasks (unless the source of culture is irreplaceable... to be 100% sure. Before testing for mycoplasma, the cells should be in continuous culture for at least...

  19. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...particular features of your plasmid, it is advisable to check out the genotype of your E. coli to ensure it is... larger culture volume can improve plasmid yield. Our lab often doubles the volume of culture used for...by adding chloramphenicol to the culture medium. Improve the culture media Changing to a richer media ...the given volume.  Let oxygen in! Make sure your culture is getting the right amount of oxygen for optimal...prevent your cultures from reaching the desired density. The volume of your flask or culture tube should...four times greater than your total culture volume and you should ensure the shaker speed is fast enough ...sufficient gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around 220...

  20. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...your incubation temperature. Some proteins aggregate at 95 °C, a common lysis temperature, so consider a...background noise. Temperature and time You may need to optimize your antibody incubation temperature and time....incubations at both temperatures if needed. Washes Check your antibody manufacturer for wash recommendations... for either a shorter (for less exposure) or longer (for more exposure) time. If you’re seeing high background...expose them. You’ll need to make sure the sensitivity of your exposure solution matches the expected amount... variants Check literature Posttranslational modifications Check literature Highly charged...Underexposed Increase exposure time Chemilumenescence: use a more sensitive exposure kit Too much ...
Showing: 101 - 120 of 1035 results