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We narrowed to 872 results for: cel

Showing: 371 - 380 of 872 results
  1. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    Blog Post
    ... expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown... complex. Not performed in a native cell context. For AP-MS, cells are lysed before the bait protein is...and interacting proteins in mammalian cells. The Journal of cell biology. PubMed PMID: 22412018. PubMed...against PPIs that aren't stable outside the native cell context. The Split-BioID method Enter Split-BioID...tagging of proteins in both BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein...proteins that interact with the bait in a native cell contact. Other PPI methods may require protein interactions...interactions to remain stable (or to be stabilized) during cell lysis and lysate fractionation. Important considerations...
  2. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    Blog Post
    ... them into your cells, you can simply look for blue colonies to determine which cells have taken up the... that the cellular fitness cost varied greatly between chromoproteins. In some cases, cells containing... mScarlet as the donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you...cloning experiment worked just by looking at the cells on the plate. For example, if your plasmid contains...Here, the chromoprotein is expressed and gives the cell a magenta (amilCP) or purple (tsPurple) color. Though...added which induces expression of the toxin. Any cells that grow after this point will be those that have...resonance energy transfer (FLIM-FRET) in living cells (Figure 4). They created ShadowR by modifying the...
  3. Plasmids 101: Optimizing Plasmid Yields

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    Blog Post
    ...large amounts of carbohydrates that can inhibit the cell lysis process. To ensure that lysis is complete,...strains adapted for protein purification use the cell’s resources to produce high levels of protein, not...difference? In the simplest of terms, more volume = more cells = more plasmid. Plasmids can vary in copy number...of plasmid copies supported within each bacterial cell in your culture. Copy number is partially specified...richer media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield... supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific...glucose (in moderation) may also serve to increase cell density and plasmid yield. Start with a single, ...
  4. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially if you...reporters allow you to easily screen or select for cells that are expressing your gene of interest at high...promoter, multicistronic plasmids ensure that any cells that are positive for your marker should also be...anytime you want to express multiple genes in the same cell. Although it is possible to drive co-expression ...promotes translation of the viral genome in eukaryotic cells.1,2 Since then, a variety of IRES sequences have...discovered - many from viruses, but also some from cellular mRNAs. What they all have in common is the ability... cistron due to factors such as the experimental cell type and the specific genes cloned into the vector...
  5. What's New in CRISPR - March 2020

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    Blog Post
    ...for studying the bacterial cell cycle, asymmetric cell division, and cellular differentiation, but it takes...crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-based DNA demethylation Optogenetic repressors...complex remains in the cytoplasm. Read the paper in Cellular and Molecular Bioengineering Find the optogenetic...Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image...lab repressed ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and...piggyBac transposon system for human pluripotent stem cells Lindy Barrett’s lab developed new fluorescently ... of multiplexed gRNAs in human pluripotent stem cells. Previously, CRISPRi or CRISPRa depended on continued...
  6. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ...synthesized inside of what is called the “target cell” (or host cell) by the viral protein reverse transcriptase...reverse-transcribed DNA into the host cell’s genome and then use cell machinery for transcription and translation...1971). Noninfectious ARK mouse embryo cell lines in which each cell has the capacity to be activated to ..., interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses are RNA-containing...separate set of instructions in the same producer cells as the virus. Because it is made at the same time...viral genes. It is derived from the host/producer cell membrane, and acquired as an envelope during exocytotic...particles do once they come in contact with target cells (Fig. 3).     Figure 3: The life cycle of...
  7. DNA Purification Without a Kit

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    Blog Post
    ...broken out of cells with a solution of salts, detergent, and ball bearings. Then a cellulose dipstick is... kits and typically follow the same principles: cells are lysed, RNAs and proteins are removed to leave... could be digested with restriction enzymes.  Cellulose nucleic acid purification Starting material: plant... plant, microbe, blood, or mammalian cells or tissue Product: total nucleic acids While glass or silica...silica are commonly used to bind and purify DNA, cellulose also efficiently binds, or at least entraps nucleic...and RNA from plants, microbes, blood or mammalian cells in 30 seconds or less without using a pipette.  ... absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids, so placing the...
  8. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    Blog Post
    ...choose cell lines that are highly transfectable.  The amounts of each plasmid transfected into cells are ... polymerase gene or using cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes...capable of being replicated and transcribed in the cell when the appropriate viral proteins are provided... RNA-dependent RNA polymerase into the infected cells. The EBOV polymerase replicates the viral genome...on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome. These...transcribed (and the viral mRNAs are translated) by cellular machinery. NP encapsidates the negative sense,...polyadenylate the mRNA. These mRNAs are translated by cellular host machinery (as viral mRNAs would be in the...
  9. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

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    Blog Post
    ... are filled with early mitotic cells as well as necrotic or dead cells (Jiang and White-Cooper, 2003)....Ivics et al 1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene called "...the fly actomyosin cytoskeleton that regulates cellular contraction. This network is composed of actin...interact with each other to generate forces that cause cells and muscles to contract depending on the context...context. Contraction is critical for processes such as cell movement/motility, development, cytokinesis, and...scientists who discovered comr thought the sperm cells in mutant flies looked like a “whole bunch of cookie...are part of a genetic cascade that determines the cell fate of the dorsal mesoderm in developing Drosophila...
  10. Plasmids 101: Stringent Regulation of Replication

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    Blog Post
    ...replicating too quickly can slow cell growth or even kill the host cell. For stringently controlled plasmids...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...be synchronized with the replication of the host cell's chromosomal DNA.  Plasmids are said to be under... stringent control when they depend on the host cell’s initiation proteins to start their own replication... occurs at the appropriate time—for example, the cell must have enough nutrients available to complete...the amount of active DnaA available is one of a cell’s best tools to regulate DNA replication. DnaA negatively...energy source, ATP binding is one more way for the cell to control DNA replication. A variety of other proteins...
Showing: 371 - 380 of 872 results