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We narrowed to 948 results for: ache

Showing: 141 - 160 of 948 results

  1. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...the cells have detached from the plate and can be isolated. When trypsinized, detached cells will exhibit...magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) Trypsin EDTA (TrypLE, Thermo...the transgene, and the specific conditions used. Each 96-well plate described in this protocol could, ...on the A549 cells described in our sample data , each 96-well plate gave rise to 15–20 monoclonal lines...cells this is about 2 × 10 6 cells in a 10 cm dish. Each 10 cm dish should be seeded in 10 mL DMEM complete...Prepare approximately 10 mL of this cell solution for each 96-well plate you plan to seed. If you choose not... Transfer 100 µL of the 5 cells/mL solution into each well of a 96-well plate. By doing this, you are ...

  2. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ...Celano of Propel Careers and Rachel Casseus, Esq. Founder of Casseus Law. Rachel Casseus Esq., Founder Casseus...guest bloggers Lauren Celano of Propel Careers and Rachel Casseus, Esq. Founder of Casseus Law. Disclaimer...numerical quota of 85,000 total H-1B visas approved each fiscal year: 65,000 visas plus 20,000 reserved for...and the arts, and requiring the attainment of a bachelor's degree or its equivalent as a minimum (8 U.S....). The foreign worker must possess at least a bachelor's degree or its equivalent and state licensure,...should only focus on your current scientific achievements. Participating in the scientific review process...international recognition for your outstanding achievements in your specific academic field, (2) that you...

  3. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...community, with approximately 100 new CRISPR depositors each year.    Figure 1: Addgene CRISPR distribution...CRISPR tools people were depositing and requesting each year (Figure 2). We did a similar analysis with ...function, by year. (B) Number of requests per month for each CRISPR function from 2012 to 2023; data are smoothed...libraries deposited and requested grows steadily each year. (The dip in 2020 is likely due to the pandemic... style When we plotted CRISPR plasmids requested each year by year of deposit (Figure 3), we saw that ... plasmids (by age) are requested and shipped out each year.     Figure 3: Percentage of requests...plasmids requested, categorized by year of deposit, for each year from 2012 to 2023.  What does this tell...

  4. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...be needed to achieve your intended next career stage and create a timeline for achieving these goals. ...and teach classes. Therefore, seek out management and conflict resolution workshops. Get teaching experience...the position, you may also want some evidence of teaching experience. How do you get there and more practically...you constructive feedback on your research, and teaches you how to write as well as mentor. Yet, even the...not always available for mentees due to travel, teaching, grant writing, etc. Also, it is unlikely that...perspectives, a shared communication style. You “get” each other. The other factor is how available the potential...writings. 8. Learn to mentor, manage, communicate, and teach The dirty secret about becoming a group leader is...

  5. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...microscopy (FLIM) method. Acceptor photobleaching method The acceptor photobleaching method is simple but limited...An EAMNET Teaching Module http://www.embl.de/eamnet/downloads/modules/FRET_teaching_module.pdf Day RN, ... knowing if these students know or interact with each other. FRET reduced the scale of our perception ...spectrum overlap. If you choose a pair too close to each other in the spectrum, you can easily directly excite...contrary, if you choose a pair that are too far from each other, you don’t have any cross-talk but you also...experiment. You can then choose the most efficient approach for your own system. The simplest and the most...fluorophores does exist in the real world and corrective approaches and appropriate controls are required to make...

  6. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ... µL of 4X sample buffer to each sample. Add 2 µL 10X reducing agent to each sample. Spin the sample briefly...peaks representing each protein band. Use the line tool to connect the bottom of each peak. Select the ...µL of 0.25 mg/mL with 10 µL PBS Transfer 5 µL of each standard to a microfuge tube. Add 8 µL of PBS to...gel. Carefully remove the comb from the gel. Rinse each well with 200 µL 1X MOPS running buffer. Fill the...skip one lane before loading samples. Load 20 µL of each recombinant antibody sample to the appropriate well...gel easier. Cover the electrophoresis chamber and attach to a power supply. Run the gel at 150 V for 1 h...Select Image . Select Type . Choose 8-bit . Determine each lane of the gel as follows: Figure 2 Using the box...

  7. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...the first three wells of each row, using a different standard solution for each row of a 96-well ELISA ...multichannel pipette to add 200 µL wash buffer to each well. Washing can be done by hand but if available...multichannel pipette, add 200 µL of blocking buffer to each well. Cover the plate with a plate seal and incubate...multichannel pipette, add 100 µL of secondary antibody to each well. Cover the plate with a plate seal and incubate...

  8. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...endogenous molecular target, meaning the dye will be attached to the target through binding of the molecular...encoded tag is required. These dyes can also be attached to an antibody to detect various targets, such...or glycans. Fluorophores can also be covalently attached to self-labeling proteins. Some common examples...covalently reacts with a small substrate ligand motif attached to a fluorophore. When paired with a protein of...higher specificity, increased versatility in probe attachment, and compatibility with far-red and near-infrared... must be taken to assess if proper staining is achieved. Fluorescent ligands have more flexibility due...peak wavelength can also decrease the rate of photobleaching. Interaction with receptors: Understanding ...

  9. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ..., but you’ll only spend half of each day on it. The other half of each day you can spend accomplishing...how much time I have blocked off for each project and how useful each item is. For instance, if I don’t ....e. those things that will be most useful for achieving your goals. However, you should have a good mixture...experiment, but you’ll have to spend the entirety of each day doing the experiment. The second way, it will...fails, you’ll still have made small accomplishments each day that should help keep you content and give you...thinking about when or how long it would take to achieve these goals. I have far too many projects to operate...Knowing this, I hand write to-do lists at the start of each day to remind myself of the specific activities ...

  10. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the desired...for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter. But...multiplexing is to clone the oligonucleotides specifying each gRNA target sequence into distinct expression vectors... using the enzyme BbsI. These expression vectors each contain Type IIS restriction sites flanking the ...but they’re optimized for different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows...generate four unique kanamycin-resistant plasmids, each containing a different gRNA target sequence downstream...you’ll clone in a polyT-termination sequence for each unused promoter. This step is necessary to generate...

  11. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...DNA-binding proteins to label certain loci, but this approach is not scalable for every locus...unlike CRISPR...chromosome-specific loci in the U20S cancer cell line./ Each color represents a gRNA targeting a specific chromosome...distances from 2-75 Mbp. Labeling multiple, intrachromosomal loci Building on the work of Chen et al., ... to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein...The specificity of these orthologs is key: since each ortholog requires a different PAM sequence, a gRNA...turned their attention to mapping pairs of intrachromosomal loci. The technique successfully resolved ... this work represents the first mapping of intrachromosomal loci, a major benchmark in characterizing ...

  12. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ... final wake it all settles down and the value of each item changes from what the reviewers so painstakingly...subjectively ascribed relative to the basic needs of each individual; the  community as a whole has yet to...and it means that there is no one-size-fits-all approach. You will have to select a tool based on your ...opinion. I could spend a lot of time running through each tool in detail, but that is already covered in the...the tools into the individual features that make each tool unique. The CRISPR Software Matchmaker is composed...organisms: This section lists the genomes supported by each tool. The genomes are denoted by organism name and...CRISPR experiments evaluate the predictive power of each tool they are interested in by comparing their past...

  13. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...transcription machinery.   Well, I’ve got some good news: complete DNA demethylation can be achieved by dCas9...can be achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved using...simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ... activity. They interact with transcriptional machinery to modify local gene expression, confounding any...and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...eLife.34870.  Kaas GA, Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013...

  14. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ... At the end of the run, each droplet is read as positive or negative for each reaction, math happens (...and departments will have qPCR machines available for use. ddPCR machines are not as commonly available...molecules added to the mastermix that are read after each cycle. The raw data is then normalized to a reference...droplet immersion technique, courtesy of CAR15.) Each droplet contains a primer and probe set for both...can be confidently interpreted as integers. Since each sample is partitioned into thousands of reactions...experiments, a number of factors will determine which approach is right for your experiment. You’ll have to consider.... PMID: 21554069. Haridan, U. S., Mokhtar, U., Machado, L. R., Abdul Aziz, A. T., Shueb, R. H., Zaid, ...

  15. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...’s lab at the University of Tokyo and Charles Gersbach’s lab at Duke University developed similar systems...light-activated CRISPR/Cas9 effector (LACE) system, the Gersbach lab utilized a similar strategy to develop an ...fusing a photoinducible, heterodimerizing domain to each of the Cas9 fragments, the authors created a photoactive...and scattered by the skin, so it is not able to reach deeper tissues. Additionally, high intensity exposure...photoswitches Photoactivation of Cas9 can also be achieved using single-chain fusion proteins by fusing Cas9...saCas9) proteins, showing the versatility of this approach. This study generated the first optically active...control over Cas9 activity. Dominik Niopek’s lab achieved this by fusing the anti-CRISPR Acr protein, AcrIIA4...

  16. Celebrating Accomplishments in the Lab

    Type
    Blog Post
    ...going away parties and potlucks for each other. The Church Lab enjoys each other’s company outside of the ...productive and successful research lab. Once goals are reached, how do science labs celebrate accomplishments ...work-life balance. How do you celebrate your lab’s achievements? Tom Ellis highlights the importance of acknowledging...to eat out together. “We also have a lab day trip each year and as we do a lot of yeast research, a few...Although hard work and perseverance are required to achieve success in the lab, it is important to take the...along the way. Take time to celebrate the lab’s achievements and enjoy time with friends and family. And ...fun outside the lab! How does your lab celebrate each other's accomplishments? Any tips on making time...

  17. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...standard curve has at least five dilutions, with each step of the series diluted by the same factor. For...example, Figure 1 depicts a 2-fold dilution series. Each step of the series dilutes by 1:2 for a series that...is diluted in a series ranging from 1:2 to 1:32. Each individual step of the series is consistent at 1...preparing a dilution series, use a new pipette tip for each step and mix the samples well by vortexing or inversion...mix thoroughly by inversion or vortexing between each step of the series. To increase the accuracy of ...  Finally, standard curves must be validated for each run. To validate, include a positive control of ... plot. For the BCA example, the concentration of each serial dilution is plotted on the x-axis and its...

  18. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...serviceman in the U.S. Army, and hockey coach all shaped his teaching style, with the former offering excellent...interpreted this trend as evidence that the best teachers and mentors pass on more than just technical knowledge...University of San Diego), decided to take a different approach. His own most impactful learning experiences happened...opportunities for his students. But he also found that his teaching responsibilities left him with limited time for...while reminding participants of the value of this approach and encouraging them to engage in the problem-...encourages faculty mentors.)  Collaboration Maximizes Reach Many new professors are wary of the time and effort...in setting up a CURE, especially if it’s a new approach within their department. So in 2016, Provost partnered...

  19. MXS Chaining

    Type
    Blog Post
    ...same cloning procedure can be used to incorporate each element into the final construct. This strategy ...Four individual expression constructs were created, each containing a unique fluorescent protein with minimal...minimal spectral overlap between them (Table 1). In each construct, 3 copies of the fluorescent protein were...enabled labeling of particular structures (Table 1). Each construct was flanked with a CMV promoter (to drive...labeling, with strong expression and detection of each fluorophore at the respective subcellular structure... 2) are contained in individual plasmids and are each flanked by the same multiple cloning site (MCS)....pattern of the MCS is regenerated at the flanks of each ligation product, which is the basis for the repeatability...

  20. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...classical TANGO assay, Kroeze et al. developed a new approach termed Parallel Receptor-ome Expression and Screening...to test multiple expression systems before they achieve the appropriate expression level for their gene...signaling pathway and includes 180 unique transcripts. Each gene was chosen to represent the primary splice ...fly genes with at least three independent gRNAs, each chosen strategically to maximize the likelihood ...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction enzyme...limited, as prior optimizations are required for each individual target species. Indeed, in order to use...system for targeted DNA methylation The traditional approach to studying epigenetic modifications has been ...
Showing: 141 - 160 of 948 results