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We narrowed to 813 results for: tes

Showing: 121 - 140 of 813 results

  1. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    Published
    Sept. 23, 2014, 3:05 p.m.
    ...scientists using pLKO.1 variants learn about those updates? This is a problem many scientists struggle with...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial...is the creation of ZappyLabs. Currently in beta testing, the software aims to be a life science protocol...Publishing Group. Like the previously mentioned sites, NPE allows scientists to join labs or groups and...Protocol Online is one of the early protocol sharing websites. It was created in 1999 and allows scientists ...protocols or provide links to protocols on their own websites. We're not sure how frequently it's being updated... With Quartzy you can share protcols with your labmates, but the system does not appear to have a way ...

  2. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...dye-nitrilotriacetic acid (NTA) conjugates (Kapanidis, Ebright, & Ebright, 2001). These Cy/NTA conjugates work the same ... What are the proper controls needed to pressure-test your FRET assay to make sure it is working? This...fluorophores to the different biomolecules you’d like to test for interaction. In fact, targeting a FRET fluorophore...attached fluorophore. However, since tRNA suppression competes with natural chain termination at stop codons,... only in a few cell types (E. coli and Xenopus oocytes, mainly). Nevertheless, tRNA suppression plasmids...tag from 6 to 10 residues. While these Cy/NTA conjugates are not commercially available, they are easily... the donor and the acceptor fluorophore binding sites are adjacent to each other on the same protein and...

  3. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ... of Antonello Bonci, Director of the National Institutes of Drug Abuse Intramural Research Program (NIH-NIDA... I work in), an obvious behavioral experiment to test for sufficiency of your circuit of interest in promoting..., on the other hand, would not be an appropriate test.  Of course, proper controls are required and include...sutures and subsequent determination of the proper coordinates of your brain region of interest (as shown in...identical for all of these surgeries. Identify coordinates through a rodent brain atlas or research papers...prior to using the virus. You may need to adjust coordinates based on your own method of identifying bregma...readout. The advantage is you can “zero” your coordinates at bregma which allows you to quickly and reliably...

  4. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...regularly, and mix samples very well.  Use duplicates or triplicates to increase the accuracy of your experiment...viral vector production facility, food allergy testing lab, and the grad student down the hall from you...-fold dilution series. Each step of the series dilutes by 1:2 for a series that ranges from 1:2–1:32. ...standard dilution series in duplicate or triplicate. Replicates allows you to calculate the curve’s coefficient...replicate measurement divided by the mean of all the replicates multiplied by 100. To calculate the %CV of the...ideal %CV is less than 15%. You should also run replicates for all of the samples in the assay, as this ...these.  Review previous literature or results from labmates to get a sense of the concentrations you might...

  5. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...restriction sites flanking the promoter-gRNA construct, but with different sequences adjacent to the sites. When...genome by removing the sequence between two target sites. Modifying multiple genes at once. Using multiplexed...promoter-gRNA constructs in order to target multiple sites! In this post, we'll cover Cas9 multiplexing options...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs. When these...unique overhangs (here, O1-4) adjacent to the cut sites “link” fragments together and drive ordered assembly...transcribed together and are flanked by specific sites that allow them to be cleaved and released. These...smaller than constructs with multiple promoter-gRNA cassettes, making them advantageous for small capacity vectors...

  6. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...CNS cell types, including microglia, astrocytes, and oligodendrocytes, provided that reporter genes specific...activators to induce gene transcription at genomic sites targeted by specific sgRNAs. Its RNA-guided nature...packaging limits (i.e., how many sgRNA expression cassettes will fit). Cre-dependent expression of the SPH...selectively target these different transcriptional start sites in order to better understand isoform-specific gene...activity (hence, “immediate-early”) and consequently regulates neuronal physiology by promoting the development...gene possesses 9 distinct transcriptional start sites, all of which yield the same BDNF protein. To demonstrate...cell types (microglia are a notable exception) necessitates such molecular gymnastics for sensitive applications...

  7. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...called a Kirby-Bauer disk diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least...shows how to perform a Kirby-Bauer disk diffusion test and check bacteria for antibiotic resistance. Source...bacteria are causing the infection before they can test for resistance. Growing and isolating the infecting...infecting bacteria can take 24 hours or more, and testing for resistance often takes an additional 18-24 hours...available. For routine infections, like strep throat, testing for antibiotic resistance isn’t commonly performed...infections is so rare. When antibiotic resistance testing is requested clinically, this involves growing ... treated. We can prescribe antibiotics for the shortest time period possible to effectively manage a patient...

  8. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ... (A) Example Level 0 assembly. BpiI recognition sites and four-base overhangs flank a coding sequence ...advisable to remove unwanted type IIS restriction sites within your sequence to improve assembly efficiency...synonymous mutation of a codon. You can domesticate the sites yourself using PCR, but gene synthesis can save ...time. Using gene synthesis, you can request the sites to be domesticated through the process, and request...sequence with the appropriate flanking restriction sites and overhangs already added. This way your synthesized...Addgene will most likely arrive in your laboratory as plates of frozen glycerol stocks. It is very important...Rob Hurt for high-throughput minipreps in 96-well plates. Plate wisely You will likely see fewer numbers...

  9. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ... bacterial ARG from Serratia sp. 39006. We next tested bARGSer with several inducible expression systems.../L yeast extract, and 5 g/L NaCl) or on LB-agar plates containing glucose and L-arabinose at 37oC for ... arabinose and colonies on arabinose-containing plates will be visibly white, and gas vesicles in cells...from left to right: colony whiteness on inducer plates, white inclusions in cells with GVs via phase contrast...mARGs also had a large genetic footprint (two cassettes + a booster cassette), their expression heterogeneity...mARGs come as a set of two PiggyBac integratable cassettes, both with doxycycline inducible promoters (Fig...increased ultrasound contrast. In all the cell lines we tested, 1 µg/mL doxycycline was sufficient to saturate...

  10. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    Published
    Jan. 23, 2020, 2:45 p.m.
    ... order fulfilment from an average of 40 minutes to 10 minutes. While users can’t tangibly see these improvements...each project. This creates accountability on the part of each developer and creates trust amongst the ...collaborate with the Product Team who in turn collaborates with stakeholders to resolve the questions.  ...would be staying on top of any third party library updates. Third party libraries are reusable software components...with their third party libraries and any security updates. Most of the time, updating these libraries is ...this year, which means there will be no further updates, security or otherwise. This might sound like Greek...

  11. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    Published
    Sept. 16, 2025, 1:15 p.m.
    ...the team tested Cas9 mutations that imposed a preference for an NGC PAM. Although all the tested mutations...whirlwind of developing cell and mouse model systems, testing base editing variants, and assessing the safety... variants (ABE8.8, ABE8.20, and ABE8e) would be tested for their base-editing efficacy in cellular models...that could target KJ’s mutation in cellular models tested both the SpG and SpRY variants of Cas9. With many...Though near-PAMless Cas9s can broadly target many sites in the genome, they also have a higher risk of off-target...bystander adenine bases were edited in cellular tests, in addition to the target base. However, all bystander... delivery components could be rigorously safety tested and reused, only the RNA portion targeting base...

  12. Tips for Getting a Faculty Position

    Type
    Blog Post
    Published
    July 20, 2023, 1:15 p.m.
    ...personal litmus test was whether my wife could explain what I was proposing. The result of my test was humbling... your candidacy. A major reason that a lot of candidates either get no interviews, or get several interviews...accomplish. No matter how many times I say this, candidates regularly try to convince me that their search...language as possible. Seriously. My suggested litmus test is whether your friend in a lab outside of your ... and therapies." This is only the first few minutes of the chalk talk. Realize that it is rare to make...available here for free!) contains strategies, hints, anecdotes, and insights. After I wrote the first version... Jennifer Lippincott-Schwartz at the National Institutes of Health, was an Associate Professor at Albert...

  13. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    Published
    June 30, 2015, 2 p.m.
    ...DNA with a TA dinucleotide (there are many such sites in the human genome). The original plasmid is empty... then degraded by the cell.4.) The transposase creates a double strand break in the DNA and allows the...has been designed to perform gene transfer in vertebrates. The system is comprised of 2 components: 1) ...number of extinct transposon sequences and then testing various combinations of changes. The basic functional.... The vectors each contain two SfiI restriction sites with different overhangs depending upon whether ... Feb 4. doi: 10.1002/biot.201400821. PubMed. 2. Mates L, et al. Molecular evolution of a novel hyperactive...transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-61. Epub 2009 ...

  14. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ... incubation (10–20 minutes) at 70 °C, or a truly lengthy incubation (30–60 minutes) at 37 °C, if you suspect...DNA-binding proteins, you may need to sonicate your lysates in order to release the proteins from their binding...in your background, try asking more experienced labmates or looking it up online. You’ll find many resources...your timer to ding. If you’re shaving off a few minutes here and there, and seeing high background in your...antibodies). You’ll want to run a protein gradient to test the antibody at the recommended dilution. If the...concentration decreases).     Figure 3: Testing the antibody’s dynamic range. A) The protein dilutions...from 1:100–1:1,000. If your antibodies come from ascites fluid, try a dilution range from 1:1,000–1:100,...

  15. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ...growth and DNA yield. Test multiple, small colonies After streaking LB agar plates, you may notice that...vectors (LTRs and ITRs), plasmids with multiple polyA sites, and other plasmids that contain repeating elements...how to amplify recombination-prone plasmids and testing your prep can save you the headache of troubleshooting...cultures contain the recombined vector backbone. Test multiple colonies before saving a glycerol stock...you can try re-streaking for single colonies and testing multiple preps. In some cases, we suggest gel extraction...the recombined plasmid is still visible. This indicates that the DNA prep contains a mixture of the full...

  16. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 27, 2016, 2:30 p.m.
    ...your incubation time on ice to 5 minutes (incubating the full 30 minutes will not significantly improve ...standard primers (no need to add unique restriction sites on the ends) and amplify your sequence of interest...the PCR product and TOPO Vector.  3. Incubate 5 Minutes at Room Temperature: You can place your reaction...digest, or sequencing. Pro tips Do not add 5’ phosphates to your PCR primers; you need that free hydroxyl...functionality are used in place of Taq to reduce error rates; however, proofreading polymerases will also remove...); however, you may need to incubate for 20-30 minutes if your PCR product is at a low concentration or...

  17. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...for the targeted gene. After initial testing, Dickinson et al. tested this system across 7 genes in parallel...remove after injection, they added flanking loxP sites and a heat-shock inducible Cre to generate a self-excising...worms have laid eggs, hygromycin is added to the plates to select for candidate knock-ins. These candidate... is inserted at the 5’ end of a gene, the SEC separates the targeted gene from its promoter, creating ...protein in the C. elegans genome. Although they tested fluorescent protein fusions, this system could ...editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Paix A, Wang Y, Smith...

  18. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    Published
    May 6, 2014, 1:07 p.m.
    .... It’s a problem that he considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch...inside cells that are on the move, such as T lymphocytes. Griesbeck and his colleagues began by isolating...version of the protein with as few calcium-binding sites as possible for added sensitivity. As reported in...these Twitch sensor variants were optimized by testing various linker configurations in a large-scale ...and high resolution functional tracking of T lymphocytes, too, making them a versatile tool for application...other people start using it. For us, it’s like a test.” References Thestrup et al. Optimized ratiometric... functional in vivo imaging of neurons and T lymphocytes. Nat Methods. 2014 Jan 5. doi: 10.1038/nmeth.2773...

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

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...research tools. BenchSci collates information about antibodies while FPbase curates information about fluorescent...sometimes formulating new biological concepts. However, rates of reproducibility for preclinical studies, as reported...Project: Cancer Biology have produced similarly low rates of reproducibility. Failed attempts to apply irreproducible... to future work cost both time and money with estimates reaching in the billions of dollars (Freedman ...making it easier to retain searchable, thorough notes on how lab procedures were actually performed, potentially... The publication record will then show that the tested hypothesis is correct (a great success story) even...category include performing experiments with too few replicates leading to suspect statistical significance (...

  20. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...reinhardtii. When exposed to blue light, this channel activates neuronal activity by opening and allowing positively...conformational change in the extracellular domain which activates the native signaling cascade of the intracellular...conformational change when exposed to blue-light and activates downstream secondary signaling pathways. Photoswitchable...light, the ligand engages the binding pocket and activates the receptor. Created with BioRender.com. ...factors will help you determine which one(s) are candidates for your experiments.  Wavelengths Most vectors...et al., 2019) for each tool, and you may have to test and optimize the intensities of light sources used... frequencies (Zhang & Cui, 2015). You’ll need to test for these differences when designing your experiment...
Showing: 121 - 140 of 813 results