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Showing: 141 - 160 of 579 results
  1. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...or more copies of the same plasmid (Bedbrook & Ausubel, 1976). This recombination event produces a multimeric...therefore run higher on an agarose gel (Bedbrook & Ausubel, 1976). In fact, if your plasmid sample has a mix...the formation of plasmid multimers (Bedbrook & Ausubel, 1976; Fishel et al., 1981). E. coli strains such...independently increase the frequency of base substitution in human cells. J Biol Chem. 2011 Mar 25;286....1021/bi00850a028. PMID: 4878700. Bedbrook JR, Ausubel FM. Recombination between bacterial plasmids leading...
  2. A Control for All Seasons

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    Blog Post
    ...negative control is a sample or group that is not subjected to the experimental condition. In the example ...control could be a third group of mice that is subjected to the experimental protocol but does not receive...knockout.  If knockout samples are unavailable, try substituting with knockdown approaches using siRNA or shRNA...either by modifying the blocking conditions or by substituting for a more specific secondary antibody. The ...Immunoblotting and Immunohistochemistry: A Guide for Submission to the British Journal of Pharmacology.” British...
  3. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...identified subpopulation. For traditional cell culture, enriching your active RNAi subpopulation can be easily...necessarily dictates the direction and scope of the subsequent two. Consequently, I will focus on different ...gene silencing efficacy, thereby requiring a substantial time investment. Once the target mRNA is cleaved...and visual screening method was limited in two substantial ways: variable silencing efficiency and labor-intensive...
  4. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

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    ...same bottle of serum again and again for each subculture can enhance the growth of mycoplasmas. The serum...generated will enter the culture medium during subculturing processes or from the air if the cell line remains...source of the problem is the media, used for subculturing, which is available in both liquid and powdered...contamination present in the serum to be used for subculturing. Avoid using expired products or reagents that...viability of cryopreserved cells decreases with every subculture. Equipment Fumigate the laminar airflow and...
  5. Cancer and the Immune System: Deciphering the Relationship

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    Blog Post
    ...itself. Many thanks to our guest blogger Subhadra Jayaraman! Subhadra Jayaraman is currently a doctoral candidate...This post was contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University... ability to inhibit further cancer growth and subjecting patients to the risk of microbial infection. ...cytotoxic effector T cells, and targeting specific DC subsets in vivo using specific antigens to activate them...
  6. A Primer on Optogenetics: Introduction and Opsin Delivery

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    Blog Post
    ... or does not result in a reward sensation and subsequent repeated stimulation. A spatial memory task, ...activation of which results in depolarization and subsequent firing of an action potential. Halorhodopsins... of which results in efflux of chloride ions, subsequent hyperpolarization, and inhibition of action potential...help restrict expression to a specific neuronal subtype. However, the length of promoter that can be used...identification of the bregma and lambda sutures and subsequent determination of the proper coordinates of your...
  7. Cellosaurus: A Cell Line Information Database

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    Blog Post
    ... lines when annotating a cell line Community submissions via email: [email protected] Entries include... lines is included when available, and can be submitted via email for cell lines profiled in the lab. ...available information about a cell line you’ve used? Submit your manuscript or data to Cellosaurus via cellosaurus...sib.swiss.This community resource is actively encouraging submissions from scientists all over globe, with some surprising...
  8. Synthetic Photobiology: Optogenetics for E. coli

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    Blog Post
    ... biological processes. Chemical effectors are suboptimal for a number of reasons, including their potential..., promoting transcription (see figure below). Subsequent exposure to red light rapidly deactivates the...computational problem of edge detection. Tabor et al. subsequently engineered a second photosensitive TCS activated...phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red light rapidly deactivates the...
  9. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

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    ... Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...to studying membranes, organelles, and other subcellular compartments is via proteomics--the study of ...promiscuous labeling enzyme and a small-molecule substrate to map the spatiotemporal distribution and interaction...sequencing, revealing that they bind to a much larger subset of protein motifs than previously thought. Sidhu...
  10. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...your experiment. To make an FMO sample, you take a subset of your experimental samples, combine it into one...be from the same host species, have the same Ig subclass, and be in the same fluorophore (e.g., if you ...staining for anti-mouse Ly6C in APC with the IgG2c subclass, you will use the APC IgG2c isotype control). ...a FMO sample: you make a sample from a combined subset of your experimental samples, stain it with every...
  11. To Each HIS Own

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    Blog Post
    ...empirically determined. In addition, some metal ion substrates, such as those containing Co2+, have a lower ...affinity for polyhistidine than more commonly used substrates such as Ni2+ and are therefore more selective... systems, consider using these more selective substrates along with stringent wash conditions.   Find ...tightly the protein will bind to the metal ion substrate. If your protein of interest is expressed at a...
  12. 5 Steps to Writing Grant Proposals for Grad Students

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    Blog Post
    ...Applications (RFA), what next? First, check the submission deadline and try not to panic if it's due in ...in mind that universities require application submission in-house approximately 2 weeks before the RFA..., award information, eligibility information, submission information, selection criteria, and award administration...written grant application builds upon the last submission. Thus, continuing to apply for funding improves...
  13. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

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    Blog Post
    ... molecule, such as biotin. Next, a conjugated substrate is added to the wells. In the case where the detection...enzyme horseradish peroxidase (HRP). Finally, the substrate of HRP is added to the wells. This reaction produces...detected by an enzyme-conjugated antibody. Next, its substrate is added and the reaction can be visualized via...that results from the reaction of HRP with its substrate, 3,3′,5,5′-Tetramethylbenzidine (TMB). The addition...
  14. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...Cas6. Regardless of the subtype, the mechanism of Cascade binding and subsequent DNA degradation appears... These type Is can then be further broken into subtypes of A–G, which come from different species and ...and type IV with an unknown effector. Type I has subtypes A through G, with different compositions of the...
  15. Of Myc and Men

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    Blog Post
    ...larger than the substance P peptide, but still quite small. However, unlike the substance P tag, they found...experiment involved a peptide from the neuropeptide substance P. This protein had a highly specific antibody...meaning the tag could be quite small. However, substance P has an amide group added to its C-terminus post-translationally...
  16. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    ...biotinylated target protein can be recovered for the subsequent steps. Phage display mRNA from the enriched pool...transcribed into cDNA fragments that are efficiently subcloned into the phagemid pDX_init plasmid, using the ...sybody pools from the phage display screens are subcloned (using FX cloning) into the expression vector ...for identification. All unique sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for...
  17. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

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    Blog Post
    ...BiFC studies used the method to interrogate the subcellular localization of interactions between basic leucine...fusion proteins it requires can lead to aberrant subcellular localization of the studied PIPs. The straightforward...endogenous counterparts using western blotting. The subcellular localization of BiFC constructs should also be...because their expression level, stability, and subcellular localization may differ from those of your BiFC...
  18. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ...synthesizing fluorophore fusions on their own. Subsequent advances in fluorescent proteins have replicated...from a sample. Chemical fluorophores can be substantially brighter and more photostable than fluorescent...reacts specifically and irreversibly with a small substrate ligand motif attached to a fluorophore (Figure...incorporation of four-membered azetidine rings could substantially enhance the brightness and photostability of...
  19. Using AAV for Neuronal Tracing

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    .... In the specific example of episodic memory, a subset of hippocampal neurons projects axons directly ...tracing has the added benefit of only labelling the subset of infected cells that also express Cre. Another...the ability to spread trans-synaptically on to subsequent neurons. AAV vectors are replication incompetent...incompetent and cannot readily be passed on to subsequent cells via replication like RABV or HSV. There are,...transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal transport. J Cell Biol...
  20. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    ...produce substances toxic to themselves? What are toxin-antitoxin systems? These toxic substances are part...inferes with DNA replication by binding the GyrA subunit of DNA gyrase and trapping it in the cleaved complex... in vitro without antibiotic selection. In a subcutaneous cancer model, the researchers showed that after...
Showing: 141 - 160 of 579 results