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Showing: 201 - 220 of 1074 results

  1. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ...what information is necessary and what can be left out, which can be a big challenge!  The resolution Throughout...glimpse into the world of science communication and can use what you've learned to share your science with...over again until you don’t.” As every researcher can attest to, science is nothing if not solving your...navigating conflict. In literary terms, conflict can be defined as (1) the struggle between two opposing... forward - and this drive is so powerful that it can entice an audience to come along on a journey even...or even by introducing a person that the audience can relate to. It doesn’t need to be anything fancy; ... provide enough information so that the audience can understand the conflict. A science-based conflict...

  2. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...….I can sense it….sound like a plot to an biology scifi movie? Or maybe it’s a biosensor?! You can do ...molecules What type of small molecules can you monitor with biosensors? You can keep tabs on my personal favorite... ‘sense’ biological compounds and reactions; you can quantify them with biosensors! In this blog we will...research are very sophisticated, but at their core they can all be boiled down pretty simply – they are systems...So now that we know what makes up a biosensor, we can discuss what flavors of sensors exist and are ready-to-use...physics, in this blog we will focus on those which can be applied in cell culture and animal models primarily...magnesium, potassium, and zinc. These ion sensors can be targeted to different cellular locations or tissues...

  3. Plasmids 101: Degron Tags

    Type
    Blog Post
    ... protein tag which can be added to either the N or C terminal of a protein and can inducibly facilitate...Protein degradation can be induced by the addition of a small molecule, and protein levels can return to normal... concentration can yield intermediate protein abundance. Finally, some degron tags can do more than just...you can’t predict with absolute certainty how a degron will behave with a new protein, but you can make... Here we will review what advantages degron tags can offer over other protein control methods and compare...addition of an exogenous small molecule). Degron tags can range in size from several kDas up to ~35 kDa and... unlike many other methods. Secondly, these tags can be used to manipulate endogenously and exogenously...

  4. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    ...understanding the scientific process can help us understand how new therapies can be developed and applied. The... it can be tempting to try to plan out a very distinct and certain path, embracing uncertainty can help...for undergrads, or anything else you can think of to ask! You can DM me on Twitter @EleanorWang. I will...too. You can follow her on Twitter @EleanorWang.     References and resources References Canning EA, LaCosse...the first step into the world of academic research can be quite daunting. During my first year of college...continue my scientific endeavors. I hope that this post can provide some perspective and advice to any undergraduate...strange and exciting revelation. Science in any field can generally be applied to improve our lives and solve...

  5. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...for recombineering can be found in Thomason et al and Sharan et al (a few strains can also be found at ...Colony PCR can be used to screen for positive clones in most cases, and restriction enzyme digest can be used...biggest limitations is that sequence modifications can only be made at restriction enzyme cut sites. The... lambda red system is an alternative method that can be used for cloning or genome engineering and is ... recombination-mediated genetic engineering.  It can be used to make an assortment of modifications: insertion...a few base pairs.  dsDNA Substrate dsDNA inserts can be made by PCR using primers that amplify the DNA...introduced during PCR. ssDNA Substrate ssDNA substrates can be generated by ordering synthetic oligonucleotides...

  6. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...3 you can see how I gated out my lymphocyte population from a tumor sample. Many cell types can be found...used a fluorescent dye that can only enter dead cells, while living cells can protect themselves from it...causes it to scatter (FSC/SSC), the detected photons can be represented through a graph of photocurrent vs... under the curve (A).   While flow cytometry can be used for virtually all kinds of experiments and... processing prior to running my flow analysis, I can however clearly identify my lymphocyte population...The voltage describes an electric potential that can be applied to the photomultiplier inside the cytometer...acquisition voltage (not shown) used in my flow analysis I can spot my lymphocyte population distinct from cellular...

  7. Five Popular Model Organisms

    Type
    Blog Post
    ...Pray, 2008). Scientific discoveries in yeast can then can be further studied in other model organisms ...transparent. Thus scientists can easily observe early developmental stages which can be difficult in other vertebrate...set of mutations that made cancer cells more resistant to chemotherapy. You can find these plasmids at Addgene...reproduce as often as every three weeks so scientists can simultaneously observe several generations of mice...organisms is that they are genetically tractable. Mice can be easily manipulated with tools like CRISPR to make...straightforward to manipulate genetically. Scientists can make a new line of flies in only 6 weeks compared...expression is possible in other model systems but can be quite difficult and time consuming. GAL4/UAS was...

  8. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...academic researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from...highlighting just a few of the many useful kits you can order from the repository: FX Cloning Kit BIOFAB...information about what these kits contain, how they can be used, and how they’ve been used to date . If you...encoding small leader peptides) downstream of which you can clone your gene of interest. Translation of the standardized...bicistronic design (BCD). The 49 vectors in this kit can be used to drive production of your favorite gene...characterized in Cambray et al., 2013. These terminators can be used to precisely tune your polycistronic E. coli... the epitope tags. Different plasmids in the kit can be used to purify proteins from E. coli, Bacillus...

  9. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...Protein can then be purified from the cellular debris. There are several popular DNA vectors that can be used...first be transcribed into messenger RNA (mRNA). mRNA can then be translated at the ribosome into polypeptide...biotechnological, and medical applications. Researches can visualize proteins in vivo by tagging them with fluorescent... structure, interactions and functions. Proteins can also be purified for use in molecular biology research...medicine (e.g. insulin). Proteins, unlike DNA which can be relatively easily synthesized, must be produced...modify your protein of interest. Modified proteins can then be extracted from lysed cells through the use... expression post-transfection. Mammalian systems can express proteins both transiently or through stable...

  10. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...panels, as fluorophores that cannot be distinguished using conventional flow can (sometimes) be distinguished...However, emission spectra of certain fluorophores can “spill over” into detectors that are not the primary...the emission spectra of the fluorescent molecules can be quite broad, detectors will usually only pick ...rainbow box” plots are unique to spectral flow and can be thought of as a series of histograms flipped onto...and R4 channels. These unique peaks in the spectra can be used to distinguish these two fluorophores from...markers that needs to be used with spectral flow—you can run a three color panel or a 40 color panel! When...level of fluorescence called autofluorescence (you can see this in your unstained flow controls). Spectral...

  11. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...line that can be transfected at high efficiency. Lentiviral transduction Cas9 and gRNA can be present... For CRISPR experiments, the following framework can help get you started: Determine what type of outcome...reporter protein such as GFP? All of these outcomes can be most effectively achieved with different CRISPR...mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator or repressor...domains to control gene expression. Base editors can help you make precise point mutations near the gRNA...components based on your model system. CRISPR reagents can be delivered via transfection, nucleofection, viral... have identified the best expression system, you can then choose the best method for introducing those...

  12. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...experiment can shed light on the mechanism by which cells gain resistance to drugs and can identify future...specific gRNAs can lead to skewed results. Selecting a cell type - Theoretically, any cell type can be used ...Identifying which genes are important for a phenotype can lead to a wealth of additional experiments investigating...sequence within the gRNA is defined by the user, and it can be easily modified to target Cas9 to virtually any...loss-of-function mutation within the target gene. CRISPR can also be used to activate or repress target genes ...genes involved in a particular phenotype. CRISPR can be readily scaled up for genome-wide screening due...-to-use lentiviral preparation from Addgene, you can skip the steps above! Cells are then transduced with...

  13. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...our quality control process for incoming plasmids can vary based on the contents of a plasmid, the first...translated amino acid sequence from within Snapgene, we can confirm the identity of the gene and check for any...any mutations. Once we identify the gene, we can easily annotate it by making a custom feature in Snapgene...multiple sequences at the same time. This means we can easily compare and analyze any differences between...plasmids are difficult to sequence by NGS. These can include GC-rich and repetitive regions. These regions...require additional verification experiments that can be partially designed using the Snapgene desktop ...Snapgene’s “Import primers from a list” function, we can detect all of the primers we have designed that will...

  14. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...out as drugs can remain present until cleared by the body. Administering the drug once can result in several... high degree of temporal resolution as receptors can be turned on and off simply with the presence or ... is immediate, on the scale of milliseconds, and can precisely link a neuron's activation to behaviors...manipulation Both optogenetic and chemogenetic tools can have their expression targeted to specific tissues...broad tissue or system-wide targeting, as the ligand can reach target cells throughout the body. Controlling...better control over these factors as the light source can be easily and quickly manipulated or turned off. ...chemogenetics, while different concentrations of ligands can be injected, this is not as precise and the offset...

  15. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...from the event can be found below along with a description of the program beneath it. You can jump to the...replicates leading to suspect statistical significance (you can read more about this problem in Button et...preparation for my talk and at the minisymposium. You can find a variety of additional resources and information...mean that someone has replicated it precisely. You can find a great discussion of different types of replication... we’ll call a study reproducible if its findings can be predictably applied to future work. Importantly...solutions based on what I’ve learned. I hope you can apply some of these solutions to your own research...vectors. When working with a plasmid from Addgene, you can be confident that you’re using the actual tool that...

  16. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...customer problems. Technical support can be highly rewarding but can also be a good jumping off point for...obvious at rather late stages (i.e. postdoc level) and can include the pressure to publish in high-impact journals...there that require exactly what life science PhDs can offer: vast practical lab expertise, experience in...may have to answer technical enquiries such as: “Can I use this product under conditions different from...might be involved in such a role. In addition, it can be advantageous to speak multiple languages - that... For particularly tricky cases, team discussions can often lead to otherwise unobtainable solutions. To...essential for success. For possibly this same reason, it can be one of the more constant and safe corners in a...

  17. Lab to Office Culture Shock

    Type
    Blog Post
    ...tight-knit team like that can and will be a little gut-wrenching, but you can get that feeling of camaraderie...I'm looking to see if there are any resources that can help with the transition.” The lab-to-desk-job culture...have as well. Making the transition from the bench can be jarring for the unprepared, and unfortunately,...academic research lab involves a few key features that can be difficult to replicate in other work settings:...experimental dry spells. Losing that sense of discovery can be hard since it’s frequently why many people get...most enjoyable aspect of their chosen career. You can confidently call yourself a Scientist. You work in...getting used to. Just remember: change is good. You can experience a real onboarding process. Unless you ...

  18. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ... homologous regions can be encoded in the primer sequences. Multiple plasmids can be included in a single...epitope tag can be moved to the N-terminus of a protein-coding sequence, or a mutation can be introduced...However, while not widely known, plasmid assembly can be achieved in vivo using a bacterial recombination...reagents or purification kits, it is low cost and can be immediately adopted by any laboratory. All procedures... Fragment generation - Any linear DNA fragments can be assembled in vivo as long as they have homologous...Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification, from...whole vector. The new codon and homologous region can be included at the 5’ end of either primer. Subcloning...

  19. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...simple presentation deceive you; pooled libraries can be quite challenging to use! In this post, I’ll walk...  When you buy a pooled library from Addgene, you can either order a DNA pooled library, which contains...lab, there are custom viral prep services that can prepare the libraries for you.  Starting from plasmids...verification, and lentiviral production steps before you can start transducing your virus. Let’s start there! ...functional titer is determined, larger lentiviral preps can be used for screening-scale transduction of cells...mutated by the viral pooled library. These cells can then be used for genetic screening, followed by sequencing... don’t. The size and composition of your library can drastically impact what conditions it needs to properly...

  20. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...network is to be always thinking “How can I help?” and not “What can you do for me?” One of the easiest ... connections can connect you to people at that organization. This type of connection can help you gather...interviews.  Pro tip: You can learn a lot from the company LinkedIn page. This can include news, inclusion...messaging, not everyone likes to. A direct email can be the best way to get in touch if you have business...t really know, follow instead of connecting. You can see what they post and occasionally comment on their... people, this option is in the “More…” menu. You can also follow companies and organizations to see their...fair game for connecting to others.  Pro tip: You can change the option for sending LinkedIn messages to...
Showing: 201 - 220 of 1074 results