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We narrowed to 1,018 results for: REN;

Showing: 241 - 260 of 1018 results

  1. Donations from Addgene to Yield Answers for Rare Disease Researchers

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    Blog Post
    ...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies...will help them to understand how different mutations lead to differences in the symptoms experienced by ...with the condition to reveal some “outstanding differences.” They now have some theories on the underlying...Institute and Addgene will make a substantial difference in pushing the work forward. Xeroderma Pigmentosum...lot of those plasmids so that we can try many different things in parallel.” Alternating Hemiplegia of...and ATP1B3 plasmids to explore the functional differences among mutations responsible for the disease known...Childhood (AHC). Patients affected by AHC suffer from recurrent attacks of unilateral paralysis (hemiplegia) that...

  2. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ... C improved assay reliability. Different primers can have different annealing efficiencies (Wang et ...of samples from different sources should not be compared. If using AAV from different sources in the same...solution, and generally these methods measure different characteristics of the solution. These characteristics...because each sample with be quantified with a different primer pair. Ct values vary based on the amount...and low-concentration samples can also behave differently in PCR, even if both samples fall within the ...reliable AAV titering? For our viral service, we currently titrate AAV vector preparations by qPCR. To ensure...absolute quantification by using the universal AAV Reference Standard Material (AAV SFM), which is an AAV sample...

  3. Important Considerations in Optogenetics Behavioral Experiments

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    Blog Post
    ...intracranial self-stimulation (ICSS) and place preference. The lab I work in (the Kreek lab) focuses on...behaviors to test reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting ...nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement but ...halves that are distinguished visually through different patterns and the animal only receives stimulation...Comparison of c-fos amongst animals that receive different types of stimulation may help to refine and optimize...many known behaviors. However, if you observe no difference in your behavioral output with optogenetic manipulation...manipulation, this can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is...

  4. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...components expressed from different plasmid backbones. They noted that backbone differences such as the order,...assess how different lentiviral plasmid backbone elements impact viral titer and to compare different Cas enzymes...Hurt for his expertise and input.  References and resources References Weber, Ernst, et al. “A Modular ...product in an affordable way, researchers test many different combinations of promoters, terminators, solubility...experimental goals are more general in nature, or you aren’t planning on using K. phaffii, then the MoClo YTK...1: A chart with MoClo kit recommendations for different modal organisms and applications.    Plant...be used to test a variety of Cas enzymes with different PAM requirements and editing abilities, such as...

  5. Antibodies 101: Flow Cytometry

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    Blog Post
    ...experiment uses 2 or 3 different fluorescent colors each measuring a different target. Flow cytometers...analysis. Since different populations of immune cells express unique Clusters of Differentiation cell surface...flow cytometry experiments.       References and resources References: Bonner WA, Hulett HR, Sweet RG, ...each cell as it passes through and scatters in different directions. The degree of scatter is captured ...fluorescent signal that is captured by detectors. The strength of the emitted photons varies from cell to cell...highly versatile and some can accommodate up to 30 different colors. This lends flow cytometry particularly...to complex experiments looking at a variety of different targets. If you are planning on using fluorescent...

  6. Fluorescent Proteins 101: Luciferases

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    Blog Post
    ...luciferases have been identified in the sea pansy (Renilla reniformis), click beetle (Pyrophorus plagiophthalamus...codon-optimized 562 61 Luciferin ATP No Sea pansy (Renilla reniformis) Natural 481 36 Coelenterazine None No ...by luciferase, releasing light. Created with BioRender.com. Bioluminescent reporters require a chemical...sensitivity. Another advantage: luciferase reporters aren’t susceptible to photobleaching — they’ll keep emitting...Cofactors/substrates Required Not required Signal strength Lower Higher Sensitivity Higher Lower Background...cruciata), and more. These variant proteins offer different kinetics, substrate requirements, and photon emission... variety of fluorescent proteins that produce different colors or have other advantageous properties. ...

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

    Type
    Blog Post
    ...Created with biorender.com. Pro tip! If your samples and control underwent different treatment processes...Created with biorender.com. The plot above is for a smeary expression and is quite different from the ...presence or absence, the plots can look quite different. You might have high expression, low expression...shows the population of interest. Created with biorender.com.   In such cases, instead of trying to quantify...yes/no flow cytometry readout. Created with biorender.com.   Using histogram plots to quantify smeary... the mean fluorescence intensity (MFI) of the different populations. MFI is the average brightness from...plot. However, the histogram plots look a bit different than they do for yes-no readouts.      ...

  8. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    Blog Post
    ...101 eBook, 1st edition.  References and resources References Swift S, Lorens J, Achacoso P, Nolan GP. ...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...from lentiviruses? There are a few important differences between engineered γ-retroviruses and lentiviruses...lentiviruses. First, these viruses were derived from different genomes (MoMLV and MSCV for γ-retrovirus; HIV ...be useful for the transduction of terminally differentiated (non-dividing) cells. Both lentiviruses and... in gene therapy research to reduce promoter interference and/or drive high expression of a transgene ...

  9. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ...nucleus, a different fix/perm kit is required (to be precise, the fixing solution is different while the...decrease in signal strength in a flow plot. d = days post treatment. Created using biorender.com. Cell viability...lineage markers, like many of the cluster of differentiation (CD) proteins found on immune cells. However...antibody host species interactions. Created using biorender.com. Another way of indirect labeling is through...primary antibody host species, as it has to be different from the other antibodies in your panel. Otherwise... dendritic cell, CD8+ T cell. Created using biorender.com.   Intracellular labeling Oftentimes, a ...first fixed and then permeabilized, using two different buffer solutions. Fixing causes protein cross-...

  10. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...graph of photocurrent vs. time. The resulting histograms is described through maximum current (H), the...cytometer to increase the electric current and thereby the signal strength. So, we need to adjust the voltage...Gates are the parameters the machine uses to differentiate between variations in those factors. To understand...a little bit of theoretical background on the different parameters that your gating strategy will be based...measured: height (H), area (A), and width (W). The different measures are represented as FSC-A/SSC-A, FSC-H...(see Figure 2). H describes the maximum signal strength, while W results from the time that the cell spent...cells, blood cells and more), each coming with a different size and structural complexity. After several ...

  11. Site Directed Mutagenesis by PCR

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    Blog Post
    ...sequence critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced...that amplifies the entire plasmid template. The parent template is removed using a methylation-dependent...A*” (in addition to the “A” site already in the parent vector). Also shown are the “B” restriction sites...with enzyme “A” as compared to the products of parental plasmid digestion (blue). Digestion with enzyme...mutagenesis. In addition, you may want to try different concentrations of template (e.g. 0.1-1.0 ng/μl...methylation-dependent enzyme (DpnI) to eliminate the parent plasmid from the PCR products, the plasmid template...patch up the DNA. The resistance marker from the parental plasmid provides a mean for selecting for transformants...

  12. Antibodies 101: Selecting the Right Antibody

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    Blog Post
    ...conjugated to different fluorophores or a panel of primaries that was raised in different species or are...are complex and antibodies may perform differently in different samples and experimental conditions. Be...for IHC, not all IHC experiments are the same. Differences in sample processing, fixation, antigen-retrieval...make sure that you can easily distinguish each different type of antibody and therefore each target protein...unique isotype or species, then you can choose different fluorophore-conjugated secondary antibodies specific...product. Note that many antibodies are sold under different names through multiple vendors, so look at epitope...to choose between the same antibody sold under different names.  If all else is equal, feel free to make...

  13. How Exercise Purges Stress and Keeps Depression at Bay

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    Blog Post
    ...expression of kynurenine aminotransferases, to enhance the conversion of kynurenine into kynurenic acid. That...skeletal muscles led to another clue: differences in the kynurenine pathway of tryptophan degradation. It...That’s key because, unlike kynurenine, kynurenic acid can’t cross the blood-brain barrier. By lowering...plasmids from Addgene. References: Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates...lowering circulating levels of kynurenine in the body, the brain is protected from stress-induced biochemical...

  14. Finding Your Perfect Job After University

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    Blog Post
    ...’s of working in different sectors. Luckily, I have worked for a variety of different organizations, so...overview of my experience and how I felt about the different positions to enable them to decide for themselves...effectively. I had to learn how to cope with a different climate and culture, and also needed to be resourceful... experience and a second language in Spanish or French, and so they were ideal candidates for these positions...very quickly; as soon as I learned how to do the different tests and became efficient, I then started to ...economy and plenty of jobs in science at world renowned institutes. The lab at the University of Queensland...diagnostic tool for the detection and diagnosis of different types of pancreatic cancer and highlighting which...

  15. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ... and reference on how to design PCR primers.  Types of PCR Since the invention of PCR, different PCR methods...Analysis RNA detection     References and Resources References Shen C-H (2019) Amplification of... methods have been developed for different scientific applications. These PCR methods all use the same...referred to as simply PCR, is used for an array of different molecular biology techniques and experiments. ...alleles or mutations. ddPCR also does not require a reference or standard curve, which can be time consuming...original template DNA in the sample.  Addgene currently uses ddPCR for AAV titrating. You can learn all...Multiple template PCR - multiple templates with different primer pairs that align to the target region of...

  16. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...genetically modify primary cells. Amongst the different vectors used, lentiviral vector constructs have...CaCl2, cationic liposome) or electrical (electric current forming membrane pores) transfection. These techniques...transducing certain quiescent or terminally differentiated cells such as macrophages and microglia. This...ability to infect non-dividing cells. Thus three different generations of lentiviral vectors have been established...transcribe the viral genome in producing cells. Currently, the third-generation lentiviral vector system...to RCL. Regarding transduction efficacy, no differences between the 2nd and the 3rd generation of vectors...marker can be placed under the control of two different promoters. Both the single and dual promoter transfer...

  17. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...about your different experiences in research labs. What are some similarities and differences you have ...found that research institutes in different places have very different cultures. In Basel, I felt that ...developing drugs against intractable diseases. Currently, a number of labs are using enChIP, and I'm very... a wide variety of biological systems. We are currently developing several additional systems of enChIP...recently developed the oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) method to suppress PCR amplification...world? Fujii: Fortunately, I was able to work in different parts of the world in my early research career...which is not trivial, and the grant situation is currently much better than that in the US for me. In addition...

  18. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...insensitivity to other ions In BRET, A mutant Renilla luciferase, catalyzes the oxidation of its substrate...applications. The system can be readily modified to use different antibiotic resistances, restriction enzymes and...either without a tag or as multiple fusions with different N- and C-terminal tags. Additionally, by using... a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with ...instance, the intra-nuclear distance between loci on different chromosomes. The advantage of having this kind...targeted loci during the cell cycle or cell differentiation. This tool should delight anyone interested...function. In order to dissect protein traffic in different systems, Franck Perez’s lab, from the Institut...

  19. Understanding GPCR Signaling Using TRUPATH

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    Blog Post
    ...number of different experimental approaches. They first assessed the activation of different members of...response to different agonists. These studies showed the promiscuity of GPCRs for different G protein transducers...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002...a vast number of signaling cascades, inducing different cellular responses. One of the biggest advantages...measure the physical association of combinations of different Gα, Gβ, and Gγ subunits. When inactive, these ...is used to excite an acceptor fluorophore. The difference for BRET is that the energy transfer is between... the signaling through the transducerome from different GPCRs. This collection can be used to study endogenous...

  20. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

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    Blog Post
    ... pivotal role in spontaneous swimming. References Arrenberg, Aristides B., Filippo Del Bene, and Herwig...Optogenetics in a transparent animal: circuit function in the larval zebrafish." Current opinion in neurobiology...wavelengths and light intensity requirements of different opsins, studies have shown that some channels,...simultaneously and still be individually activated with different lights (Baier et al., 2009).  Zebrafish behaviors... do so, the lab expressed the LiGluR opsin in different subsets of spinal cord neurons using the Gal4/...Gal4/UAS system. This created fish lines with different groups of neurons that could be activated with light...spontaneous swim or escape, was elicited when these different neuron groups were activated or “turned on.” In...
Showing: 241 - 260 of 1018 results