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We narrowed to 1,075 results for: CAN

Showing: 221 - 240 of 1075 results

  1. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...packaging vectors for larger genes - they can hold up to 8 kb, while AAV can only carry around 4.5 kb. Different...Behaviors you can study using viral vectors By modulating neuronal gene expression and activity, you can use viral...development of viral vectors, behavioral neuroscientists can now precisely control where and when certain genes...required, because seemingly tiny external factors can have a quite noticeable influence on the outcome ... several days preceding the experiment. Handling can however negatively interfere with certain tests and...and affect behavior. A fact to consider: Rodents can hear ultrasonic frequencies (> 20 kHz, produced from... left behind from a preceding animal (ex: urine) can be distracting. If it is not cleaned, animals tested...

  2. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...measure how many of the viral particles produced can actually infect cells. These assays typically involve...several commercially available p24 ELISA kits that can quantify p24 levels in a matter of hours. A major...quantification tend to be overestimates since they can include free p24 and defective viral particles. Direct...backbones; once validated these universal primers can be used to titer any lentiviral prep that shares ...p24 assay, titering via measurement of viral RNA can overestimate the amount of infectious virus in a ...defective particles. Studies have found that values can be 10-1000 fold higher than for functional assays...from amplification of the original plasmid which can over-inflate results. A packing vector deficient ...

  3. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...fluorophores, and can bind and light up epitope tagged proteins in vivo. These probes however can be expensive...optimized elements of an antibody probe, scientists can now more easily visualize single molecules and newly...protein sequences called epitopes and scientists can use these to tag their proteins of interest. Antibody...experiments demonstrated that the HA frankenbody can be used to amplify fluorescent signals in in vivo...effective in sensitive cells such as living neurons, and can be fused to other fluorescent proteins and tags including... to get started with your experiments! What else can you do with the HA frankenbody? Another large advantage...minutes in living cells, meaning that scientists can track the movement and localization of single HA-...

  4. Viral Vectors 101: Transductions

    Type
    Blog Post
    ... transductions can offer benefits such as tissue-specificity, high efficiency, and can be selected to ...which can’t be easily achieved with alternative methods. Ready, set, transduce! Making virus can sound... or AAV. Then, your desired DNA to be integrated can be cloned into the vector that you have selected....resulting in functional virions. These viral particles can infect other cells, but since the majority of the...production cell line (depending on the virus), virus can be collected for the transduction step. Viral particles...viral collection fairly easy. Producer cell media can be collected, centrifuged to remove any producer ...rogue HEK-293 cell). This virus-containing media can then be placed on whatever cells you would like to...

  5. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...technique can be used as a preliminary validation on your bulk population. This way you can ensure you... you can determine if it is worthwhile to isolate individual clones from that population. You can additionally...such as CRISPResso can help with the data analysis. If NGS is not an option, you can still detect off-target...systems generate a deletion large enough that it can be visualized by running your PCR on a gel. It’s ...sure your PCR amplicon isn’t too large so that you can easily visualize the size shift you anticipate. A... a new recognition sequence. In these cases, you can screen using PCR and a restriction enzyme digest....endogenous restriction enzyme sites? Never fear! You can introduce a silent, passenger edit to serve as a ...

  6. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...Twitter because I can get a little opinionated and, while it can be anonymous, I can always block the ... (short for “Ask Me Anything”) on r/Science. You can see some of Joanne’s comments on the AMA process ...Reddit is a volunteer-organized platform where users can participate in forum discussions about a wide variety...scientific fields. They also host AMA forums where users can ask designated scientists anything about a particular...active community of people interested in science. You can see some of the conversations going on within the...place for us. We’re definitely glad we did - you can see some of the stats from the AMA to the right -...The AMA process was simple and well organized. You can check out the r/Science AMA Submission Guidelines...

  7. Bringing Sustainable Practices to the Lab: Easy Action Items

    Type
    Blog Post
    ...problem, how can one little change make a difference? Individual changes to our behavior can lead to cultural...where I’ll list some easy but effective habits you can incorporate to reduce your lab consumption and increase...posts. Making small changes in your daily habits can feel ineffective or unimportant. With so many big...cultural shifts that can have bigger impacts. For example, if you joined an office where everyone used ...cups. By initiating little changes in ourselves, we can both influence others to change their behavior and... the lab Here are a few easy and fast things you can start doing right away to reduce your consumption...makes a big difference. Clean out freezers so you can make room and don’t have to keep buying more freezers...

  8. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...proteins is that the tag can alter the structure of the protein. In some cases this can lead to changes or ...staining can be beneficial when expression levels are low as multiple secondary antibodies can bind to ...protocols? If this sounds like you, then flow cytometry can help you overcome these challenges and make these...When combined with cell sorters, this technology can be used to isolate a specific subset of cells in ...fusions In addition to visible light, flow cytometers can have a variety of fluorescent light sources. When...cytometers however are highly versatile and some can accommodate up to 30 different colors. This lends... In addition, the level and timing of expression can also be affected by the tag.  Protein expression,...

  9. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ... Alex Schier’s Lab. gRNAs can be in vitro transcribed from ssDNA, which can be generated by commercial...incorporated into the genome     Off-target effects can occur due to prolonged Cas9 expression     The requirement...compared to plasmid-based expression of Cas9/gRNA and can be used for most of the current genome engineering...using RNPs for CRISPR experiments. The RNP method can often be used in cells that are difficult to transfect...transfect, such as primary cells. Using RNPs can also alleviate difficulties with protein expression that...protein and a gRNA from a commercial vendor, this can often be expensive. An alternative approach is to...experiments using RNPs is the variety of methods that can be used to deliver a Cas9-gRNA RNP. This advantage...

  10. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...because organic solvents can be quite harsh - lipids and soluble proteins can be lost during fixation ...the protein, which can disrupt protein structure and thus structural epitopes, but can reveal linear epitopes...organic solvents can be used to simultaneously fix and permeabilize a sample. But you can also use organic... you might come across a couple other terms that can cause some confusion. Paraformaldehyde: Many IF ...Along with preserving your sample, cross-linking can block the epitope of some targets, preventing your... to be washed away. Furthermore, these chemicals can also alter the structure of proteins, which could... improve antibody binding for some targets, they can destroy it for others. That said, organic solvents...

  11. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ... Overexpression of fluorescently tagged proteins can lead to improper protein localization, protein aggregation...fluorescent protein fusion. CRISPR genome editing can help accomplish this goal. The power of this system... specified by a guide RNA (gRNA) sequence. Users can design gRNAs to induce the break at a specific genomic...pathway, a new user-defined DNA sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists... contain your newly tagged gene. Most insertions can only be detected by labor-intensive processes like...high-throughput targeting vectors. These vectors can be used to tag endogenous genes and simultaneously...connector and tag donor plasmids from the library can be used to generate functionally unique repair templates...

  12. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...example) and can be analyzed using Addgene’s Analyze Sequence tool. Excellent, how can I check if the... of web pages the plasmid page is nested in. You can see in the example above that the bread crumbs trail...plasmids with similar names in the collection, but you can use this name to find this plasmid from our search...out this video for a walkthrough of the ways you can search our website). A quicker way to find this plasmid...out the plasmid's structural features and see if I can easily clone my gene into it? Your best bet is ...plasmid maps as part of the deposit process and these can be found in this location as well. These maps make...provide plasmid maps whenever possible. GenBank files can also be uploaded onto our plasmid pages, but these...

  13. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...these biosensors can be found at Addgene. Split FPs – some FPs (e.g. GFP, Venus) can be split into two...Photoactivatable / Photoconvertible: These proteins can switch their color when activated by a specific excitation...wavelength. This means that the emission wavelength can change. In a few cases, the initial state of the ...proteins change their color over time. Therefore, these can be used as “timers” for cellular processes following...they will form the full FP and fluoresce. Split FPs can be used to determine the proximity of two proteins.../em peak. Therefore, choose FPs that your system can excite, and detect the emission. For example, if ...yellow light). Note that some combinations of FPs can cause an effect called FRET (fluorescence [or Förster...

  14. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ... it can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be ...are doing. But aside from sequencing, scientists can also study the microbiome by using engineered genetic...IncP⍺-family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative...plasmids (pGT) with various replicative origins that can be used for broad or narrow range of microbial species...a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated with antibiotic...Collins’s lab developed a paper-based method that can be used to detect microbes and biomarkers on paper...method is the RNA toehold switch, a RNA hairpin that can be designed to bind and detect nearly any RNA sequence...

  15. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...backbones and specific primers you can use to confirm your clones. You can also find a list of popular sequencing...gRNA insert or not. A blue-white screen, however, can give you this information (more on this below). Plating.... lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse lactose. Fortunately, for the lab researcher... will just see the vector backbone band. Or, you can choose an enzyme that cuts once in the plasmid backbone...one band. These are only a few examples of how you can use restriction digest to screen your clones, but...using restriction digests to analyze plasmids, which can help you visualize this process, or visit our restriction...restriction analysis blog post. Colony PCR Colony PCR can detect the presence or absence of DNA using lysed...

  16. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...These whorls can be quantified for individual cells, and the number of 'normal' looking cells can be counted...that are engineered can be shared to simplify the comparison by others. Together, we can build a valuable...probes. You can follow him on twitter: @joachimgoedhart.   Marieke Mastop is a PhD candidate at the Section...tendency to form higher order aggregates. This property can be tolerated in some applications (e.g. labeling ...detail here. There are a variety of methods that can be used to measure your fluorescent protein’s propensity...proteins to form homodimers or higher order oligomers can be analyzed by several techniques. Gel filtration...spectroscopic methods (based on fluorescence polarization) can be used to detect homodimerization. However, all ...

  17. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    ...that antibodies can do! For instance, some Fc receptors, once bound to an antibody, can act as immune modulators...to bind. We also know that changing the Fc domain can impact an antibody's binding affinity (Janda et al... the different immune responses? We got you! You can also review antibodies in our Antibodies 101 animation...different receptors, but their downstream effects can be sorted into three main effector functions.  ...Antibody-dependent complement deposition (ADCD) Antibodies can bind to a pathogen and physically prevent it from...effector function known as neutralization. But they can also bind to a pathogen and mark it as a target for... cell. If the pathogen is a large parasite, ADCC can be induced through a very similar interaction between...

  18. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ... insertion line can then be isolated and screened. Depending on the marker, traps can be used as high-...frame, in fact, they can be activated a considerable distance away from a gene which can make identification...expression levels (high/low) and patterns (tissue/timing) can be monitored under various conditions without necessarily...uncharacterized gene. The actual reporter construct can be something phenotypic (such as a fluorescent protein... result in lethality before all of its functions can be observed. Functionally redundant genes – multiple...multiple genes which function in overlapping roles can mask the depletion of one another. Under these circumstances...reporter gene flanked by a minimal promoter which can integrate proximal to an endogenous gene to turn ...

  19. Finding Your Science Policy Path

    Type
    Blog Post
    ...you can jump from one type of position to another throughout your career. Plus, just one role can encompass...  Where can you do science policy? Because the field of science policy is so diverse, you can do it in... of work can happen at the national level within the executive branch or in Congress, or can also happen... programs can connect you with alumni before and during your application process so you can learn more... about virtual meet ups and happy hours that you can attend to help build relationships.  Reach out to...differently depending on how and where you pursue it. It can manifest via research and data gathering, regulatory...as a profession. Figure 1: Science policy can lead you to many paths! For a more details, check...

  20. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...In the event that genome sequencing can be done, this technique can also help pinpoint the genes within...Nebulization units, which can be purchased cheaply, are small plastic containers that can easily be attached ...are hard to find Sequencing and cloning phage DNA can be difficult, in part because phages frequently alter...approximately gene-sized) fragments so that these fragments can be cloned into E. coli and subsequent proteins screened...screened for cell-binding abilities (6). This can be accomplished using a restriction enzyme with a short...DNA with phi29 polymerase prior to fragmentation can greatly facilitate downstream manipulations. Phi29...Phi29 polymerase is a phage-derived polymerase that can perform whole genome amplification (by a mechanism...
Showing: 221 - 240 of 1075 results