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We narrowed to 849 results for: NTS

Showing: 141 - 160 of 849 results

  1. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    Published
    Aug. 22, 2023, 1 p.m.
    ... Identify reagents you will need, any required training, and a priority list of experiments you hope to...that they may want to consider for future work placements. The chance to learn and collaborate all while...which you can apply to. These structured types of placements often have some form of built-in professional... ensure your expectations are reasonable, your reagents are onsite before you need them, and you are set... plan as well. Making logistical and funding arrangements Visiting scientist positions take commitment...structured visiting program, you will have to make arrangements on your own. This will include, but is not limited...transportation, living expenses, and necessary reagents. Your primary lab may be willing to cover these...

  2. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ...of your various blot reagents. There are two types of gradients. Protein gradients use dilutions of your...background. Created with biorender.com   Both gradients can be informative, so spend some time thinking...particular question before running one.  Good on gradients? Excellent! Now, let’s head west, intrepid reader...   Ladders If you need very accurate protein measurements, consider using an unstained protein ladder,...conjugated antibody. If you do not need such precise measurements, you can use a colorimetric ladder, visible ... after your transfer. If you are expecting low amounts of protein, make sure your reversible staining ...wet and semi-dry transfers, you can adjust the amounts of alcohol and SDS in your transfer buffers. To...

  3. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...chemicals (biotinylation reagents). There are a variety of biotinylation reagents available that differ ...and thus different reagents are useful for biotinylation in different microenvironments. There are also ...organisms for life. Biotin is only synthesized by plants, most bacteria and some fungi, thus mammals must...unaffected by changes in pH, temperature or organic solvents. Uses of biotin in molecular biology Biotin is...protein purification and immunoprecipitation experiments. Biotinylated proteins or complexes can be easily...immunohistochemistry and antibody labelling. In these experiments, biotin is added to a primary or secondary antibody... ideal for signal amplification during these experiments. During detection, a biotinylated reporter enzyme...

  4. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...sequencing large amounts of DNA quickly and accurately. We're sure to see many improvements in a variety ...and sequencing gels, as well as their modern equivalents and see what we can learn from their evolution...DNA identity, size, and abundance. If a scientist wants to know if an organism has a particular gene mutation...confirming the results of cloning or knockout experiments to forensics and clinical diagnostics. One example...individual without sickle cell anemia, two DNA fragments are observed because the DNA has the site where...investigate DNA sequence content by comparing the fragments created by different restriction enzyme digests...allows a researcher to properly orient specific fragments of a DNA sequence of interest to create a physical...

  5. Communicating Your Science With Help From ComSciCon

    Type
    Blog Post
    Published
    July 19, 2018, 1:12 p.m.
    ... of graduate students who are passionate about communicating science.  ComSciCon's events have 3 main ...apply for our future events. In addition to this annual event, graduate students in different regional...comscicon.com/. All our events are offered free of charge to accepted students, and we pay for travel ...Communicating Science Conference series for graduate students. I believe that communication is the single most...main goals: To help graduate students in all STEM fields become excellent science communicators. To empower...empower graduate students to become ambassadors for their respective fields. To help attendees launch ...practice science communication to more graduate students.   Each June, graduate student leaders in science...

  6. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ... methods. Across these methods, the following components are needed: γ-Retroviral transfer plasmid encoding...similar to lentiviral packaging methods. The three components described above (envelope, packaging, and transfer...because once in the host cell cytoplasm, the viral components are small enough to enter the nucleus through...slightly different isoforms of these packaging components. Therefore, lentiviruses may not be efficiently...plasmid backbone is meant to be descriptive of the components found within the backbone. For example: pLXSN...particle containing Matrix, Capsid, and Nucleocapsid components. Pol Precursor protein containing Reverse...Reverse Transcriptase and Integrase components. Transfer cPPT Central polypurine tract; recognition...

  7. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...color changing flower involves two basic components: the pigments (anthocyanins), and the plant circadian...associated intellectual property concerns – patents and licensing agreements. Developing a product without the ... the rewards are great if you take the plunge – plants are the foundation of life on earth, and opportunities...feeds, foods, and fibers. And because working with plants can be challenging, there are a lot of unexplored...the complicated and unique metabolic processes of plants are being investigated on limited funding and time...also identified and characterized mechanisms and components of the circadian rhythm in petunias. Because ...before us, it’s possible for us to combine these components to develop flowers that change color. Without...

  8. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...vacuolar) pH environments that inactivate SEP and leave only DsRed to fluoresce (1). With variants that can...facilitate innovative cancer research, the Reference Reagents Group of the NCI RAS Initiative, led by Dominic...method, which allows for easy assembly of DNA fragments by utilizing regions of sequence homology (usually...and test constructs composed of complex genetic elements. To demonstrate the usefulness of GAMP, the Jacks...promoters expressing GFP, tetracycline response elements, and shRNAs many of which are available at Addgene...allows for easy and rapid targeting of complex elements to specific genomic sites. For instance, the authors... community. If you plan to use GMAP for your experiments, think about depositing your new plasmids with...

  9. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...technology itself is constantly improving. Cas9 variants have been modified for genome editing, activating...specificity of the Cas9 nuclease with engineered variants: eSpCas9, SpCas9-HF1, & HypaCas9. The off-target...substitutions throughout the groove in 32 separate Cas9 mutants. When tested for their ability to cut the EMX1 ...embryonic kidney (HEK) cells, 11 of these initial mutants retained the on-target activity of wt Cas9 while... of these mutations and tested the combination mutants for their on-target and off-target activity at ...locations in HEK cells. Further testing of two of the mutants, SpCas9(K855A) and eSpCas9(1.1), followed by a ...double strand breaks (BLESS), revealed that these mutants do not cause off-target effects at unanticipated...

  10. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ... compatible with detergents like SDS, while BCA assays are not with denaturing agents like DTT or β-mercaptoethanol...other hand, they are not as sensitive to lower amounts of protein, and can require more antibody or loading... loading buffer will not contain any denaturing agents. However, you’ll still have to lyse your proteins...protein. Proteins located in membrane-bound subcompartments, like the mitochondria or nucleus, will likely...gradient gel, the decision is probably already made: gradients are extremely difficult to handcast, so unless...blots. If you need to preserve sample, check the ingredients before using, as some contain Tween-20, which...reaction rate and are compatible with antimicrobial agents. Chemiluminescence kits When activating your enzyme...

  11. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...structures or other local genomic elements (Hazan and Bester 2021). Other Cas variants Another option is CRISPR...ncRNA (Jiang et al. 2014).  Simple knock-out experiments are trickier when it comes to long non-coding... et al. 2018). However, removing large lncRNA fragments with multiple gRNAs poses the risk of disrupting...can be minimized by disrupting lncRNA regulatory elements that are crucial for transcription and downstream...or introduced, into the break sites. Knock-in experiments are important for visualizing ncRNA localization...long ncRNAs is impractical. Instead, tags or stop elements can be inserted to disrupt the transcription of...CRISPR/Cas9 system is not effective for your experiments, there are plenty of other CRISPR-based options...

  12. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...only will these mStayGold variants be valuable tools for your next experiments, they will also likely inspire...genome editing for primary cells Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate... Breaking up is hard to do: Monomeric StayGold variants By Mike Lacy In 2022, Atsushi Miyawaki and colleagues...stability. Here’s a quick overview of these new variants:   Figure 2: Inner mitochondrial membranes...The Miyawaki lab generated a series of StayGold mutants and identified a triple mutant, Y187F, R144I and...Addgene here!   Ando, R., et al. (2023). StayGold variants for molecular fusion and membrane-targeting applications...gene delivery capsids previously identified in rodents have not been widely successful in primates. The...

  13. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...may undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying the...contact A variety of reagents can increase transduction efficiency. These reagents typically enhance the...working with virus: 1. Viral vectors undergo DNA rearrangements. Depending on the composition of the viral...viral vector with bacteria designed to minimize rearrangements such as NEB Stable or use a standard strain...ensuring viral production is to perform PCR against components of the viral vector using the harvested viral... you time, and allow you to standardize your experiments later. Stay tuned to the Addgene blog and be ... pages to get up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A...

  14. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...to extract without harsh reagents and patience. Plant expression systems Plants provide a cheap and low-tech...through the use of protein tags and separated from contaminants using a variety of purification methods. Deciding...systems can be scaled up for production of large amounts of protein. Some drawbacks of insect cell expression... systems. Bacterial expression systems When one wants to produce vast quantities of protein rapidly and... DNA vectors that can be used to produce large amounts of protein in bacterial cells: the pET, pRSET, ...recombinant proteins. Many cells from various types of plants such as maize, tobacco, rice, sugarcane, and even...share many of the same features and processing requirements as mammalian cell expression systems, including...

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

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...Contamination by humans accounts for the largest source among those mentioned above. The contaminants can spread through...antibiotics and are broad-spectrum antibiotic agents. These agents can be used to wash the cell cultures. The...to the liquid or semi-solid medium (containing nutrients essential for mycoplasma growth). Infected cell...synthesized by eukaryotic cells. First, the kit components rupture the mycoplasma and then specific enzymes...for the lab to avoid bringing in environmental contaminants.  Figure 2: An Addgenie works in the...Photo by Kate Harten DeMaio. Cell lines and reagents Examine your cell line stock for mycoplasma contamination... subculturing. Avoid using expired products or reagents that have been stored for a longer time. Use fresh...

  16. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...repetitive genetic elements that are found in both prokaryotic and eukaryotic genomes.  Components of a transposon...around in the lab.  Let’s take a look at the key components of transposons, how they are classified, and ...target sites. Integration of most transposable elements (TEs) results in the duplication of this target...insertion.  Types of transposons Transposable elements are split into two classes based on their mechanism...the rest of this article. Class I Transposable elements (TEs): Retrotransposons  Class I TEs are also ...screens Transposons, by their nature, are mutagenic elements, which makes them a great tool for mutagenesis...developed the INTEGRATE (Insertion of Transposable Elements by Guide RNA-Assisted Targeting) system. INTEGRATE...

  17. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...DNA you’re purifying (plasmid, genomic, or DNA fragments from agarose).    In this article, we will go ...or agarose gel slices Product: plasmids or DNA fragments The plasmid purification and DNA gel extraction...Starting material: agarose gel slices Product: DNA fragments You might be wondering, “How is milk going to ...or even paper towels to purify DNA and RNA from plants, microbes, blood or mammalian cells in 30 seconds...into this mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids...placing the dipstick into a wash solution removes contaminants while nucleic acids stick around. Finally, purified...gel slices Product: purified PCR product or DNA fragments Maybe you’re not ready to give up DNA purification...

  18. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...immunity in diverse tissue environments. These studies provide quantitative measurements of cellular motility... tissues structures are more stable and local movements predominate (e.g. a basal progenitor cell migrating...proteins can and have been used to monitor cellular movements in the immune system. The techniques used here... Studying cell mobility within these complex environments has never been easy and, for many years, researchers...to count cells and statistically analyze cell movements. By using cells labelled by fluorescent chemicals...giving researchers the ability to monitor cellular movements in real time. This technique was first Introduced...use of fluorescent proteins to track cellular movements and interactions. Even if virtually all fluorescent...

  19. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    Published
    July 20, 2021, 1:15 p.m.
    ...permeabilization with detergents such as triton-x, saponin, or tween. Take care when using strong detergents like triton-x... and electronics systems, FACS instruments have specialized components that can divert a subset of cells...overcome these challenges and make these types of experiments easier and faster to perform. Introduction to...channel number based on intensity. Higher intensity events are assigned to higher channels. In a typical assay...assay a user will compare the shifting intensity of events. Since intensity positively correlates with expression... flow cytometry particularly well to complex experiments looking at a variety of different targets. If...for setting up antibody based flow cytometry experiments.       References and resources References:...

  20. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...analyzing thousands of edits performed over multiple experiments and comparing the robustness, accuracy, and speed...also provides a Knockout score (KO-score) which represents the proportion of cells that have either a frameshift... How ICE works   Following delivery of CRISPR components into target cells, genomic DNA from both edited...analysis, which can compare up to five editing experiments at a time, and batch analysis, which compares...confident you can be in the ICE score. KO Score - Represents the proportion of cells that have either a frameshift...the edited pool. The black vertical dotted line represents the cut site, and yellow + symbol on the far ...trace. The horizontal black underlined region represents the guide sequence, along with the red underlined...
Showing: 141 - 160 of 849 results