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We narrowed to 1,052 results for: sta

Showing: 181 - 200 of 1052 results

  1. Cancer and the Immune System: Deciphering the Relationship

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    Blog Post
    ...development of drug resistance in bacteria, the tumor cells that escape the elimination stage acquire genetic...favorably attaining the equilibrium stage. This can be called “stalemate equilibrium” where the cancer stops...cancers differ in practically every facet starting from manifestation to their specific interactions with different...the immune mechanisms of the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer...the acute immune responses that eliminate early stage cancer cells, chronic inflammatory responses can... tumor cells spreading to other organs during metastasis. Evading Recognition Tumor cells over-express...damage healthy cells, and often cannot eradicate metastasis or eliminate the probability of a relapse. Chemical...

  2. Polymerase Chain Reaction Overview and Applications

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    Blog Post
    ...). Taq polymerase is thus a heat resistant enzyme that can withstand changes in temperature. Taq was first...the starting template DNA the fluorescent signal throughout the reaction is compared to a standard curve...detected compared to the standard curve can be used to determine the amount of starting material in your sample...Chemistry for his work. Since then PCR has become a standard and essential practice in molecular biology and...heating and cooling steps. PCR relies on heat resistant DNA polymerase from the thermophilic bacterium...range in which the forward and reverse primers are stable. At this temperature the primers can anneal to ...stranded DNA template strands. DNA polymerase is also stable at this temperature and can bind to the primers...

  3. Plasmids 101: Secondary Nanobody Toolbox

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    Blog Post
    ...comparison of this one-step staining with nanobodies vs two-step staining with antibodies. Nanobodies...before application to the sample. Staining with this multi-color staining workflow yields similar localization...localization patterns as cells stained for one target (compare nanobodies staining in figure 4A vs 4B). How...structure, HCabs are like a pared down version of a standard IgG. See below for a comparison of nanobody, HCab...double-edged sword. An antibody’s large size creates more distance between the target antigen and the fluorescent... fusion with a reporter enzyme. For multicolor staining, careful consideration must be used when selecting...different IgG subclasses of 1° for multi-color staining Can be expensive Nanobodies   Recombinant...

  4. Mesothelioma - Causes, Symptoms, and Treatment

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    Blog Post
    ...usually started to spread (metastasize) and turned into Stage 3 or Stage 4 cancer. These later stages of a...indicate, for Stage 3, that the cancer is large in size and may have spread while Stage 4 indicates that...types of cancer have different challenges and obstacles to overcome, mesothelioma sufferers can see great...first exposure to asbestos to the time when someone starts seeing – or feeling – the first mesothelioma symptoms... time. Doctors and researchers don’t entirely understand why some people develop the disease while others...do not work very well. The best option in such instances is palliative care, which helps patients manage...discover the cancer earlier, in particular before it starts to spread beyond its point of origin. One of the...

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

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    Blog Post
    ...Fluorescent DNA staining is another DNA staining alternative. This method uses DNA stains like DAPI and ...the dry state for a longer time. When they come in contact with a nutrient source, they start proliferating...guest blogger, Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology.... The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial drugs like penicillin... human scalp, sneezing, coughing, etc. Also, a constant in-flow of individuals wherever the cell cultures...this task, you can use PCR based detection, DNA staining, fluorescent tagging, agar plating, etc. Be wise...Culturing on agar plates/broth is considered the ‘gold standard’ for detecting mycoplasmas. In this method, the...

  6. Technical Design of a Western Blot

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    Blog Post
    ...for your standards (ladder). You’ll want to select a pre-stained or unstained set of standards that covers...But if you understand the technical aspects of a western blot, you'll be able to understand how to modify...have enough lanes available for your standard curve, your standard, and your sample(s), as you can only...methods and buffers to choose from. If you are starting from tissue, you’ll likely want to use a mechanical... V, 1–2 hours Good separation, quality, and stability Limited protein size range Tris-acetate ... visualize proteins on a PVDF membrane without staining; simply wet your membrane with 20% methanol and...you’re not running a western. They are not very stable and will noticeably degrade after several months...

  7. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...enhancer sequences in different chromatin states. Alexander Stark and colleagues at the Research Institute...their position relative to a transcription start site. In STARR-seq, randomly sheared genomic libraries ...(Q3) MARCH 2016Enhancer activity mapping with STARR-seq   Although many technologies have been developed...Pathology have developed an alternative method, called STARR-seq (self-transcribing active regulatory region ...over input levels using next gen sequencing. The STARR-seq backbone is available from Addgene in both human...including but not limited to cell growth and drug resistance. Recently, the Wu Lab at Texas Tech developed...plasmid or as individual plasmids shipped as agar stabs. Iverson, et al. ACS Synth Bio. 2015. PubMed PMID...

  8. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    Blog Post
    ...produce substances toxic to themselves? What are toxin-antitoxin systems? These toxic substances are part...these systems is that the toxin is stable while the antitoxin is unstable, meaning that the cells must continually...the pair is more stable than the other. Here, the restriction endonuclease is more stable than the methyltransferase...degraded leaving the cell to be killed by the more stable toxin. Cells only survive if they retain the plasmid... contains the mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked...) contained within it by selecting for Zeocin resistance. Then, counter-selection with methanol medium... unmethylated sites in the chromosome. The more stable endonuclease can then cut the chromosome leading...

  9. Meet an Addgenie: Insights into Grad School, Career Paths, and More

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    Blog Post
    ...leave. I was really devastated when that happened. I didn't know if I should even stay in science.” ...by choice, but by circumstance. She was working as a research and technical assistant, when her life changed... all of the Meet an Addgenies features on Instagram! Staying true to your values While going through the... major and a Spanish minor, it’s time for me to start considering my future path. Talking to Addgenies... what lies ahead. Thus the “Meet an Addgenie” Instagram interview series was born. Over the six months...who has been in many roles in Addgene since he started, said, “the thing I wanted more of [after undergrad... a few years ago at a CRISPR meeting.Read his Instagram feature here. You also don’t have to go back...

  10. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ...using biorender.com.   Dye stains Dye stains are non-antibody-based stains that bind DNA or free amines...alongside standard surface labels. Here, I will introduce you to a few useful labeling and staining techniques...techniques beyond standard surface labeling. Pro tip! Many people use 'labeling' and 'staining' interchangeably... it comes to viability and proliferation staining. Dye stains are membrane-permeant or -impermeant and...nucleus, or visualizing DNA instead of proteins. Some stains can diffuse through the cell membrane freely, while...Addgene uses 'labeling' to refer to antibodies and 'staining' to refer to small molecule dyes. Indirect labeling...cross-linking, killing the cell but preserving the cellular state (think of mummification). Permeabilization, as the...

  11. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...differentiate between variations in those factors. To understand the principles of gating, you’ll first need to... the magnitude of FSC, SSC, and all fluorescent stains. The cytometer's software will allow you to adjust...specific gates (single cells, live cells, my marker stains of interest). Let’s double-click inside our gate...duplicates can’t be reliably analyzed on their staining signals. For this, you’ll need to gate based on...here, I actually want to gate on my negative, unstained population, which I can clearly differentiate ...positive one (Figure 8). Note that in many cases a stain will not result in clearly distinct positive and...in between. Again, it can depend on the type of stain whether a tighter or looser gate makes sense. Here...

  12. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    ... a small group of scientists including Stanley Falkow, Stanley Cohen, Herbert Boyer, and Charles Brinton...treated a tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102,...EPISOME-MEDIATED TRANSFER OF DRUG RESISTANCE IN ENTEROBACTERIACEAE VIII: Six-Drug-Resistance R Factor.” Journal of...the 1940s and 50s, scientists were working to understand genetic cytoplasmic factors that could be transferred...other extrachromosomal particles, particularly Resistance or R-factors. Like F-factors, R-factors could... selected for E. coli transformants that were resistant to both. When this proved successful, pSC101 became...in these cloning experiments. Although plasmids started as a somewhat niche area of research, they are ...

  13. R Bodies: Membrane-Rupturing Microscopic Tools

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    Blog Post
    ...While the importance of understanding these complex processes can’t be overstated, we can also learn a lot...solutions to transmitting forces over long distances. For instance, how much force can be generated by conformational... a structured material over a long distance? And can we understand such a structure well enough to engineer...dynamically rearranging into different polymerization states. Instead, R body monomers respond to pH changes...

  14. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ...guarantee 100% transfection efficiency. Stable transfection A stable transfection is used to create a population...genetic material directly into the genome to form a stable transfection. Unlike bacterial transformation, ...described below, are typically employed when creating stable cell lines and are not used for general plasmid...this ever-versatile fluorophore in a later post, so stay tuned! Transient transfection and the elusive "mammalian...expression in yeast and bacteria, plasmids used for stable transfections rarely contain an ORI since the integrated... subsequently pass it on to future generations. Stably transfected cells may be considered an entirely...Positive selection in mammalian cells To achieve stable transfection, there should be a selective pressure...

  15. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...functional antibiotic resistance. Colonies with an insert in pUCXKT will therefore have resistance to both kanamycin...the genome and to determine, for instance, the intra-nuclear distance between loci on different chromosomes...many paths. Understanding how a protein traffics through these pathways is key to understanding, and potentially...ingenious two-state assay based on the reversible interaction between a hook protein stably localized in...and monitor individual living cells in order to understand how they work. Light activation allows scientists...This leads to 2048 (211) possible combinations of states. Each of the integrases is supplied on its own ....1038/nmeth.3147. Bioluminescent pH sensor Understanding intracellular pH regulation is important as ...

  16. Transferable Skills Guide: Creativity

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    Blog Post
    ...your ideas are not well received, understand that people can be resistant to change. Ask your peers or your...: creativity. I’ve heard quite a few scientists state that they’re not creative, usually saying that they...creative problem-solving process?  You may want to start by carving out time to tackle it. It’s easy to get...Another strategy for your brainstorm session is to start with something completely unrelated to the issue...Originals by Adam Grant, a creativity expert at Stanford conducted an experiment in which people were asked...were instructed to use the three-ring binder as a starting point, the ideas generated were hardly novel. .... When told to use a roller skate as a starting point, their ideas scored 37% higher in originality. It...

  17. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    ...not yet have a comprehensive understanding of how cancers become resistant, an emerging theme is that they...involved in drug resistance Wood wanted to find out which particular pathways drive resistance to specific ...relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said...David Sabatini at the Whitehead Institute and MIT, started with a list of 17 signaling pathways that are frequently... a time, you could identify those that confer resistance to drugs,” in a rapid, straightforward, and scalable...Wood says, and it took them about six months from start to finish. They cloned all the cDNAs into a single... for use in screening for pathways that drive resistance to drugs in vitro, and they’ve shown that it ...

  18. Giving gRNAs a Facelift - Synthetic and Beyond

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    Blog Post
    ...worrying about the status of your RNA. gRNAs modifications are even going beyond stability, adding color and...modified) for optimal stability and efficacy.  Base modifications What is more stable than RNA? DNA! One...their precious RNA samples. RNA is inherently less stable than most macromolecules used in scientific research...or protein. It comes as no surprise then that stability of the RNA used in CRISPR-Cas9 experiments – the... and even on-off switches. Stabilizing gRNA modifications  Sugar phosphate backbone modifications The...phosphate molecules of the RNA backbone to increase stability.  Nature has evolved ways to prevent degradation... protect it from nucleolytic attack. Another stabilizing sugar modification, found at the same site, is...

  19. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    Blog Post
    ...such as Golgi staining, injection of diffusible labels, or electron microscopy of stained sections, were...characteristics (low aggregation, high photostability, and high stability after fixation.) From the seven resulting...transient Cre recombination is required to prevent constant inversion of the construct. Sanes and Lichtman...intensity, caused partially by fluorophore photoinstability, as well as the tendency of fluorophores to...Second, the system did not permit analysis by immunostaining. Although the fluorophores used are fluorescently...Brainbow-1 and Brainbow-2 each contained a “default” state; for example, Brainbow-1.0 expresses RFP when the...construct has not undergone recombination. This default state was disproportionately expressed by a majority of...

  20. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ... designed to insert a resistance gene. The vector contains a neomycin resistance gene (NeoR) flanked by...insert and later remove a resistance gene. In this construct, the neomycin resistance gene is floxed so that...into the vector on either side of an antibiotic resistance gene. To avoid selecting for cells in which the...perform a positive selection for the antibiotic resistance gene and then a counter selection for the negative...removed from the chromosome and and replaced by the resistance gene.The gene is thus disrupted, or knocked out...occurred. Neomycin, puromycin, and hygromycin resistance genes are commonly used for positive selection...positive selection marker. Since an antibiotic resistance gene can affect a cell's phenotype, researchers...
Showing: 181 - 200 of 1052 results