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Showing: 161 - 180 of 1074 results

  1. Antibodies 101: Affinity Tags

    Type
    Blog Post
    ... is a 26 kDa tag which can be isolated with glutathione resin. Like MBP, it can be removed via protease...Some of these tags can also be used for protein purification, just as affinity tags can sometimes be repurposed...With a little thoughtful planning, affinity tags can make protein purification a cinch. Types of affinity...used primarily as protein purification tools (and can have secondary functions as detection aids.) When... naturally exist in most expression systems that can be added to the N or C terminus of your protein of...Maltose binding protein (MBP) is a 45 kDa tag which can be purified (along with its fusion protein) with ...with amylose resin. The MBP tag, although large, can actually increase expression and solubility of the tagged...

  2. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ...system. The first two can serve as controls in a variety of experiments, while rtTA can be used to activate...that can be used to deliver fluorescent proteins to cells as determined by the serotype. You can use these...that you can use to map neuronal connections. Expanding the viral vector knowledge base: You can help! We...from select vectors in the repository yourself, you can now order ready-to-use virus directly from Addgene...many researchers we’ve met with have asked if we can help them share and distribute other reagents. Virus...while still ordering viral vectors. Virus production can be a frustrating process that requires a large investment...viral production and testing so that researchers can use our educational resources and experience to their...

  3. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...factors that can affect DNA stability in solution are pH and DNase contamination.  Extreme pH can denature...sample over time and determine which experiments you can effectively use the DNA. pH and DNases affect DNA...stranded DNA are repaired by base excision repair, but can lead to mutations in single stranded DNA undergoing...undergoing replication.   DNase on the other hand can digest and degrade the double helix, fragmenting plasmid...degradation, and/or the use of chelating agents that can be introduced in the purification protocol to remove...substantial degradation and which experiments you can use your plasmid.  DNA elution choices: TE, Tris ...the stability of your DNA, it will limit what you can do with these samples. This is because divalent cations...

  4. Antibodies 101: ChIP

    Type
    Blog Post
    ...tissue sample can also need significant optimization for the sample type and size.) It can be quite the...protein and DNA are no longer bound, and the DNA can then be isolated and studied through a variety of...application for exploring these interactions, and can give valuable insight into DNA expression and protein...approach for the faint of heart!  The Process ChIP can be done for either tissues or cells. The process ...most ChIP experiences, but the increased binding can mask the epitopes antibodies recognize. It is therefore...before moving on to the next step. Typically, this can be done by cutting the tissue sample up and then ...usually the preferred method, enzymatic digestion can be appropriate when your protein of interest is a...

  5. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... gradients can be informative, so spend some time thinking about which type of gradient can help answer... gels can overheat. Running your transfer in the cold room or packing the box with ice packs can help ...from the same lot as you need or can get from the manufacturer. You can also consider using monoclonal ... high, you can consider adding a detergent like Tween-20 to your washes — although this can be harsh and...Western blots can be tricky to get right. If you’ve thoughtfully done the technical design of your blot... optimization and troubleshooting.   Before you can optimize or troubleshoot a western, you’ll need to...Created with biorender.com   Alternatively, you can run a reagent gradient. In this approach, you’ll ...

  6. Early Career Researcher Toolbox: Free Tools for Science Writing

    Type
    Blog Post
    ...is stored in the cloud, so you can access it from anywhere, and it can be shared with others. The Web ...Connector, papers can be saved from Firefox or Chrome browsers with just one click. Papers can be cited in ...LibreOffice, or Google Docs. Files can be synced across devices, and you can share your library with others...calendar after completing your writing goal, which can serve as positive reinforcement for a habit. The ...technique timer  One time management technique that you can use as part of your writing schedule is the Pomodoro...is called a “Pomodoro.” After four Pomodoros, you can take a longer 15-30 minute break. The method is named...word suggestions. De-jargonizer The De-jargonizer can identify jargon in your writing. While it’s not possible...

  7. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...inactivating genes. CRISPR can target a genomic sequence and create a break that can be repaired by homologous...designed in the lab. Specially constructed plasmids can be used to replace genes in yeast, mice, or Drosophila...This final product of non-homologous recombination can survive positive selection, but it will be sensitive...selection marker. Since an antibiotic resistance gene can affect a cell's phenotype, researchers usually remove...through the insertion of flanking LoxP sequences, you can remove the gene through the addition of Cre recombinase...the neomycin resistance gene is floxed so that it can be removed post-selection through Cre-mediated recombination... (knocked in). As seen in the above example, you can remove a floxed resistance gene using Cre recombinase...

  8. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... equipment can be defined as equipment with capital costs greater than $5,000 but this can depend on the...where you can start to identify which items aren’t critical to your research success. If you can get away...loss of time, can exceed the cost of the new equipment. That’s why a resource budget can help you compare...your research, negotiations can be much easier. However, negotiations can still go horribly awry when ...resources needed to accomplish your science, and can include: equipment, reagents, animals, glassware,...emotional weight. Therefore, too much focus on money can dissuade a productive negotiation. A more productive...much those resources will cost and how these costs can be reduced. Negotiating is easier when both parties...

  9. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens can be used to inhibit... may be most effective. Once drug candidates are isolated, they can be validated for specificity to their...or their direct target can be identified if it isn’t known. If a strong candidate emerges, downstream optimization...added QC is where CRISPR can save time and money to make sure quality candidates are selected.      ... targeted design. Alternatively, a disease model can be generated, and drugs screened for a change in ... workflow? CRISPR both speeds up the process and can help weed out a lot of poor drugs before time and... affect disease progression or onset. These hits can reveal individual genes or sometimes even genetic...

  10. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts in your experiments....Medicine, to learn how problematic oligomerization can be and how to avoid it in your experiments. Better...proteins are incredibly useful, chemical fluorophores can still be used in a wide variety of experiments. Read...produce reporter proteins. These toehold switches can be used to quickly detect the presence of Zika virus...School of Medicine, this post describes a tool you can use to study basic virus biology while leaving virulent...transcription as well as reporter genes. To learn how you can use minigenomes in your research, check out this ...-bodies, “force generating protein machines that can extend and retract over many microns in response ...

  11. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...system or cell type can completely fail in a different system (Issa et al., 2023). This can be particularly..., which can lead to cells in far away brain regions expressing your gene of interest. This can cause issues...circuit can occur. Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous... chicken-beta-actin or CAG can drive strong early gene expression, but can become methylated over time...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig....Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst... validation first.  AAV-mediated gene expression can differ extensively depending on the experimental ...

  12. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ... POI can affect protein folding and protein-protein interactions. Also, overexpression of FPs can trigger...other). Combined with AAVs, FPs can be easily expressed in the brain and can be detected through their direct...protein partners within the cell. However, antibodies can have low affinity for these tags and even multimeric...epitopes tags, FPs are intrinsically fluorescent and can therefore be used in live imaging experiments crucial...bright, stable and well tolerated by cells. They can be used for protein localization, isolation, and ...that emit across the visible spectrum, researchers can also use multiple FPs in single experiments and visualize...These probes are both intrinsically fluorescent and can be detected with highly epitope-specific, fluorescently-labeled...

  13. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ...antibody can be directly conjugated with a reporter enzyme or a conjugated secondary antibody can be used...conjugated antibody. While labs can conjugate antibodies themselves, the process can be laborious and expensive...conjugation methods can interfere with binding, and some common antibody buffer components can interfere with...accuracy, specificity, and sensitivity (a good ELISA can detect picogram quantities of the target!) and use...antibodies are conjugated with a reporter or tag that can then be detected.  Typically, the reporter is an ...often opt for the chromogenic substrates since they can be read on a standard plate reader as opposed to ...saving time. Moreover, since secondary antibodies can cross-react with samples, eliminating them from the...

  14. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...  If a plate of samples is ever dropped, we can simply scan the tubes and know what is in each one. Even...samples, it can be done easily with a barcode reader and an Excel spreadsheet. Start by scanning the sample...shown below). This number ensures that every sample can be tracked easily at each step in the storage process...sample at Addgene has four layers of location which can all be extrapolated from the 2D barcode and LIMS:...Tubes are assigned a location within a plate that can hold up to 96 tubes (see image below). Each plate...metal freezer rack that contains five levels that can hold five plates on each level. This rack is located...

  15. All in a Twist: dsRNA

    Type
    Blog Post
    ...diseases can result (Fanconi Anemia, cancer, Fragile X syndrome, etc.). Hopefully now you can appreciate...of these viruses have one thing in common – they can generate dsRNA as part of their infection and/or ... inherently double stranded, while ssRNA viruses can generate dsRNA intermediates as part of the viral...circulates, but upon viral infection, this nucleic acid can build up. The foreign dsRNA activates the innate ...typically RNA that the RNAi is designed against), RISC can repress translation and/or degrade target mRNA. ...such as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription or ...epigenetically silenced, but dysregulation of these systems can lead to local dsRNA production. In the same vein,...

  16. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ... Basically, if you can devise an experimental assay that works with flies, you can utilize the resources... Seahorse can be employed to look at oxygen consumption. Drosophila are also used in cancer research (... I’ll provide a brief introduction as to how you can engineer new transgenic flies to study your favorite...others use attP sequence docking sites so that you can control exactly in which locus your transgene ends...1): Random insertion has the advantage that you can get varying expression levels of your transgene in...for genetic recombination so that the transgenes can be inherited on the same chromosome. Targeted Insertion...which chromosome your transgene has integrated. This can be accomplished with a few matings to balancer chromosome...

  17. Fostering communication in remote collaborations

    Type
    Blog Post
    ...everyone can access data and drafts of manuscripts and grant applications in real-time so they can provide...Science is a global endeavor, and that can often mean that scientists who live across the country or ... these accelerate the progress of research, they can also present hurdles for the team of researchers ...disciplines or doing different tasks. But these challenges can be exaggerated when communicating across time zones... that helped make my experience run smoothly and can help other teams successfully overcome these challenges...every month is long enough that some experiments can get done and data analyzed between each meeting, ...conducting the experiments to the PIs. Remote meetings can be tricky, and it takes extra effort to make sure...

  18. PhD Applications After COVID

    Type
    Blog Post
    ...online meeting can take some of that pressure off. This question and answer process can also help you ...campus even if you can’t visit. By talking with people at the institution, you can get a better sense ...the in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity... flood of interview requests and visitation days can be, at times, very overwhelming. As someone who applied...technologies that helped us navigate a global pandemic can also be the thing that provides a new perspective... applications opening. With virtual options, you can narrow down your list of schools by meeting with ... idea of that advisor’s research interests which can be helpful in creating your application to that program...

  19. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ...). While some mutations can remain stable as homozygous alleles, others cannot. Geneticists need a method...happens. Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome? Continue...Continue reading and I’ll show you how Drosophila can be used to push your research in new and exciting...of seeing a phenotype since additional genes that can do the same function are less likely to exist). If...system), and the third post will describe how you can make your own mutant flies. Find Drosophila Resources...~9 days after setting up the genetic cross). You can tell when pupae are ready to hatch the next day because...their bodies will be very light in coloration and can look very plump. On the side of their abdomen will...

  20. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...Both of these can be increased by concentrating a viral stock. Different titering methods can reveal differences...virus solution can be obtained. 4. Enhancing virus-cell contact A variety of reagents can increase transduction..., you can check if virus is being generated by your transfected packaging cells. This check can be performed...An estimated 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages... viruses are everywhere and doing everything, it can be annoying when we try to use them in an experiment...which are factors in the technical challenges they can present. Many researchers find themselves plagued...titers (which is unavoidable for some vectors) you can concentrate your virus by ultracentrifugation of ...
Showing: 161 - 180 of 1074 results