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Showing: 261 - 280 of 899 results

  1. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    Published
    Oct. 2, 2018, 12:56 p.m.
    ...lab alumni as you can or schedule video/calls if they are not local. Alumni meetings can be the most informative...lab such a big deal? It’s actually something you CAN choose and it will make a huge difference for your...right and responsibility to choose a lab where you can thrive and do your best work. This post is focused...choosing a lab, but almost all of these guidelines can apply to any workplace or job. It amazes me when ...questions in the sections below. Mentoring relationships can help identify your priorities and needs, but sometimes...informative way of gathering data. Former lab members can give open, revealing information about the culture...opportunities with the lab head. Former lab members can give you a fuller picture. Consider talking to people...

  2. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...so much for how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires...fluorescent proteins (FPs) can be problematic because excitation light can cause phototoxicity, autofluorescence...tools are fantastic examples of technologies that can accelerate your research. The creation of NanoLuc...by external light sources. These external sources can generate autofluorescence and have limited penetration...protein fusion (OgNLuc). Both fluorescent proteins can be excited by Nluc emission and the lab has dubbed...nm – a clear demonstration of why a BRET reporter can improve deep tissue imaging. The constructs for GpNLuc.... The technique of coupling FPs with Luciferases can overcome these problems but has been hampered by ...

  3. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ...not all cell types can be labeled, and that while multiple copies of the transgene can integrate into cells...recombination is required before fluorescence can occur. Cre recombinase can be used alongside common, existing ...label an organ or entire organism. Because each cell can have multiple copies of the transgene that will recombine...express the construct but do not undergo recombination can be detected by immunostaining of the truncated YFP...and fluoresce in the same color as the fused XFP can be used to amplify the signal. Imaging endogenous... extremely versatile technology as recombination can be induced in developing fish via Cre protein microinjection...mouse (Snippert et al., 2010), a reporter mouse that can be used to track any cell type when crossed with ...

  4. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...nuclease, can be used in conjunction with Cas13 to amplify the signal of a single target. Csm6 can cleave...laboratories. In the case of the COVID-19 pandemic, qPCR can be used to diagnose the presence of SARS-CoV-2 RNA...reporter. The detectable signal from this reporter can be measured and quantified to determine the presence...an RNA guided RNase (Abudayyeh et al 2016). Cas13 can be guided by a single CRISPR RNA (crRNA) to cleave...non-specific ssRNA cleavage. How does SHERLOCK work? Cas13 can be programmed with crRNA to target an ssRNA of interest...recognizes and binds to the programmed sequence, it can promiscuously cleave surrounding ssRNA molecules....speed, ease of use, and portability. This method can be used for an array of applications including clinical...

  5. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    Published
    Aug. 20, 2024, 1:15 p.m.
    ...may not be clearly picked up by the sensors. You can adjust this by changing the voltage for the particular...1). Based on where the GFP- population ends, you can draw a gate which selects anything to the right of...expression, instead of its presence or absence, the plots can look quite different. You might have high expression...upregulated or downregulated based on your experiment. You can see from Figure 2 that determining where to place...quantify smeary plots For smeary expressions, you can quantify shifts in signal using the mean fluorescence...peak that shifts to the left. When using MFI, you can still continue gating populations of interest with...expression to the baseline/negative control. You can see how it is easier to visualize these four populations...

  6. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...protein cannot bind. After an inducer binds to the activator protein, the activator protein can bind to...this post, we’ll discuss inducible promoters, which can be switched from an OFF to an ON state, and how you...inducible promoters regulated? Inducible promoters can be regulated by positive or negative control. Positive...temperature, and light are all examples of factors that can lead to the induction of a promoter. Below, you’ll...cAMP complexed with cAMP activator protein (CAP) can also stimulate AraC binding to I1 and I2 sites. Supplementing...near-zero expression at regular temperatures but can be induced by heat or cold exposure. Examples include... pBad is likely a better option since expression can be repressed by glucose. Temperature inducible promoters...

  7. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ...If there’s a plasmid or AAV tool that you can’t find, you can always suggest a plasmid to us, request that...including a team that created a novel form of AI that can learn new tasks and beat humans at the ancient Chinese...I met one researcher studying how the microbiome can affect Alzheimer's - a connection I wouldn’t have... get into the field, there’s likely some way you can use the tools and skills you’ve developed in your...maybe your favorite FRET-based biosensing technique can be applied to an important neuronal signalling pathway...ve got a useful tool, there’s likely someway you can apply it in neuroscience. AAV tools from Addgene ...useful, these vectors require a bit of expertise and can be difficult for individual labs to make efficiently...

  8. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ... with large inserts can make lentivirus production frustrating - a researcher can complete an entire lentiviral... SpCas9 and Cas9 nickase for genome editing. You can also enhance expression of your gene of interest ...EGFP, BRAF, and MAP3K4. The gRNAs targeting EGFP can be used as positive controls for gene editing or ...repository are CRISPR pooled gRNA libraries that can be used to screen for genes involved in a wide variety...Getting these quality-controlled reagents from Addgene can greatly expedite a researcher's ability to make discoveries...you’ll also receive transfer plasmid DNA that you can use along with lentiviral packaging plasmids to create...standard lentivirus production protocol here and you can find more tips and protocols for working with lentivirus...

  9. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...modification systems, yet can still affect whether certain restriction enzymes can effectively cleave DNA...of mutated plasmids which can later be verified by Sanger sequencing. How can I tell if my enzyme will ... of the DNA surrounding your restriction site(s) can help you quickly predict which (if any) of those ...sequence visualization software (we used Snapgene here) can help you assess whether your restriction site will...in Lane 2. Controlling methylation Finally, you can control methylation by altering your choice of bacteria...that will be blocked by Dam or Dcm methylation, you can ensure this site remains unblocked by first cloning...

  10. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...undergo nick repair. This method can work well, but poor template digestion can result in a high number of ...Cloning, and SLIC can all be used to assemble multiple fragments, but these methods can be expensive because... causation in molecular biology and genetics. It can be used to characterize the function of certain regions...biomedical research, modeling patient mutations using SDM can help determine if a variant is causal for a given...strategies, only 3-4 phage proteins are needed and these can be provided via a separate plasmid or be inserted...machinery is induced using L-arabinose. These cells can be made and frozen in large batches suitable for ...

  11. How to Lead a Great Meeting

    Type
    Blog Post
    Published
    Feb. 9, 2016, 3:30 p.m.
    ... to our “real” work. However, a well-run meeting can have quite the opposite effect. A great meeting should... meeting. Follow the tips below to learn how you can run a top-notch meeting. Define the meeting Define...some other commitment. An announcement of purpose can help everyone to ‘land’ in the meeting and get focused... direction you’d like to pursue? Undefined scope can lead to long meetings spent on details that do not... sure to keep them manageable. At Addgene, teams can add as many points as they like to the written agenda...we like to send around a Google doc where people can make comments. At meetings, we just focus on the ...meeting (see “defining the scope” above) or if you can cut it down to thirty minutes. Keep it moving: ...

  12. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    Published
    July 26, 2018, 12:46 p.m.
    ..., which can be difficult to predict. Slower Dephosphorylation Rates: FRET-based reporters can have slower...measuring kinase activity at the single-cell level can aid in drawing connections between signaling activity...spectral properties that limit which fluorophores can be multiplexed together. Challenging  to Design: ...cytoplasm and/or nucleus, while FRET-based reporters can be used to measure kinase activity in specific subcellular...and the cytoplasm, nuclear export and import rates can influence KTR translocation. Work from the Covert...

  13. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...control of DNA recombination and gene expression can be advanced by the coordinated use of fellow SSR ...asymmetric core. Recombination between these sites can lead to excision, inversion, and translocation of...Continuous inversion of the DNA sequence of interest can lead to poor expression of genes within that sequence... for Cre and FLP function. Because recombination can only occur between target sites of the same sequence...recombinase - keeping Cre out until needed But what can be done when the recombination should only occur ... cortical neurons and their synapses.   How can you label a single neuron and its synapses in the...the living brain? Cells of a specific brain area can be transfected by in utero electroporation. To achieve...

  14. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ... DNA strands - a double-strand break (DSB). DSBs can be caused by intracellular factors such as nucleases...identical to the original DNA sequence at the DSB, or it can introduce specific mutations into the damaged DNA...displaces one strand of the homologous DNA donor and can template repair off the other strand; this results... loop (D loop). The recombination intermediates can then resolve to complete the DNA repair process ...Homology directed repair pathways HDR mechanisms can be divided into two main camps: non-conservative ...presence of only one invading strand at a DSB that can be used for repair (a one-ended DSB). This single...HDR is a powerful site-specific knock-in tool that can be harnessed by researchers for many applications...

  15. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...your lentiviral vector will infect, pseudotyping can help. In pseudotyping, you produce viral vectors ...cytotoxicity and the mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein...syncytias that cause the cells to die. Cytotoxicity can also be cell type dependent: if a cell has few receptors...lentivirus (Crawford et al., 2020). That way, they can study the spike protein’s role in infecting cell ...methods.  In terms of preparation, VSV-G is stable and can withstand ultracentrifugation which helps scientists...cells and neurons. However, VSV-G’s broad tropism can be undesired for in vivo studies since there’s the...the potential for off-target infectivity as it can bind and transduce many cell types before reaching ...

  16. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...methods and machines one team can learn so linking together expert teams can push a scientific project to...too), we will often look for partnerships where we can leverage another organization’s expertise to our ...meeting people with different mindsets and approaches can truly broaden your mind. When I first began collaborating...Getting companies to come to these types of events can be very tough. First I needed to find the right contacts...guidance. The relationships you establish in this way can, of course, lead to jobs and even favors, but you...don’t forget about collaborations and projects you can do with others after work. Get sponsors for a training...accomplish with people at your organization. You can meet collaborators  for lunch, make a point of talking...

  17. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...research as well. For example, CRISPRa can be used to find potential cancer drug targets by finding genes that...these can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used... the potential to be used in therapeutics, as it can increase gene expression at certain loci to create...al., 2016). Additionally, CRISPR-based activation can be made inducible upon environmental triggers. This...This is done by creating a split dCas9 protein that can reassemble when exposed to specific UV wavelengths...under native conditions, the less activation you can achieve using a CRISPRa. Your gene of interest might... four guides directed towards that gene as there can be a large difference in guide potency. Thus, in ...

  18. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...that now can be deposited at Addgene. What makes these fluorescent proteins unique and what can they be...mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not significantly excite ...green and yellow fluorescent protein wavelengths, it can be imaged either with standard green fluorescent ...reduction in emission readout, respectively. mNeonGreen can be used as a fusion tag for a wide range of applications... other common fluorescent proteins and therefore can be used in immunoprecipitation experiments with other... mCerulean imaged with a standard CFP set. mTFP1 can be used as a FRET donor to mCitrine and mOrange or...

  19. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...2002) and many downstream complexes involved, it can be difficult for scientists to understand how GPCR...transducers (collectively known as the transducerome) that can be activated downstream of GPCRs that are not involved...by a GPCR on the cell’s surface into action that can result in the initiation of a vast number of signaling...downstream signaling pathways.  This dissociation can be measured using a technique known as Bioluminescence... a decreased GFP2 emission. The activation state can be measured by a BRET2 ratio, which is the ratio ...GFP2 emission to RLuc8 emission (see figure). What can TRUPATH be used for? The Roth lab used the TRUPATH...transducerome from different GPCRs. This collection can be used to study endogenous ligands, the pathways...

  20. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...plasmids can be used to do include: Produce large amounts of a protein so that scientists can purify and...one-stop reference guide for plasmids, so that you can spend less time researching the basics and spend ...point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids...make more of the plasmids as they grow, scientists can easily make large amounts of plasmid to manipulate... Produce proteins that glow so that scientists can track their location or quantity inside a cell ...genome engineering) Produce synthetic viruses that can be used in research or for therapeutics Addgene ...
Showing: 261 - 280 of 899 results