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We narrowed to 929 results for: KIN

Showing: 551 - 600 of 929 results

  1. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...reference protein) under identical conditions, a ranking of the practical brightness can be made (Goedhart...repeating this with another fluorescent protein, a ranking of practical brightness can be made. This strategy...their practical brightness in a system closely mimicking the future application. In figure 2 we provide...

  2. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...high-throughput manner and therefore push us closer to making cancer precision medicine a reality. Challenges...Since time is of the essence in clinical settings, looking ahead, we’re starting to build saturating libraries... Institute of Harvard and MIT and is focused on making precision functional genomics a reality for future...

  3. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...think. So, if you’ve ever wondered what goes into ‘making virus’ or what gag and pol stand for, we demystify...recombinant adenovirus is missing the endogenous E1 gene, making it replication deficient. This is typically supplied...discussed are compiled for reference below. Happy virus making!  References and Resources Kulanayake, S., Tikoo...

  4. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...can begin to understand how the brain works. It’s kind of like “the head bone’s connected to the neck bone... to the rest of the body. Therefore, by simply looking at how neurons are physically connected we can ...efficiently deliver genetic information or tools, making them useful tracers, but finding a virus that works...

  6. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...better fuels, feeds, foods, and fibers. And because working with plants can be challenging, there are a lot...construct our color changing flower we are carefully making our way through the IP landscape, using plant parts...novel pieces where applicable. We will also be working with open source, transformation technologies like...

  7. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    ... and devoted 100% of his energy and effort into making the startup self-sustainable. Within a year from...all safety nets.” Raising seed money. “Before seeking funding, ask yourself if you really want to do ...Co-founder and formerly VP Operations of Sample6) for taking the time to speak with us about the biotech startup...

  9. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...this point, you may be asking yourself, what type of Fab fragment are we talking about here? Many of the...fragment. But this bivalency may cause problems for blocking purposes, since the fragment could bind not only...

  10. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...someone speaks about an affinity reagent, they’re talking about a standard antibody: An ~150kD protein with...nanobodies, are derived from a camelid antibody lacking a light chain, called a heavy chain antibody (Muyldermans...clear: it’s an exciting time to be a scientist looking for that perfect fit!  References and Resources...

  11. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...between them, could affect the properties or trafficking of the fusion protein (Kimple et al., 2013). ...opsin-expressing cells). For other things to keep in mind when making fluorescent protein fusions, check out Joachim ...Fluorescent Proteins 101: GFP Fusion Proteins – Making the Right Connection.     Thank you to our guest...

  12. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...for individual cells, and the number of 'normal' looking cells can be counted. This assay will provide information...on the dimerization tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not ...

  13. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...density: particle concentration is measured by looking at absorbance at 260nm using UV/Vis spectrophotometry.... The purity of the sample can be estimated by looking at the ration of DNA (260nm) versus protein (280nm...calculated in terms of plaque forming units (pfu), taking initial vector dilution into account. You'...

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

    Type
    Blog Post
    ...laboratory of Dr. Moshe Szyf at McGill University, working to develop CRISPR-based tools for epigenetic editing...Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun...Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013) TET1 Controls CNS 5...

  15. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, ...articles describing the FRET tool that you are looking for and check our curated list of biosensors too... Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking the ruler to the jungle: Single-molecule FRET for...

  16. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope...whole segment—a GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into...and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing the tag...

  17. Hot Plasmids February 2024

    Type
    Blog Post
    ...10.1038/s41587-023-01756-1. PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants ...with some applications. Several labs have been working to produce monomeric versions that break up the...was delivered to brain endothelial cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when...

  18. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...Scientists looking for more compact genome editors due to delivery constraints and scientists seeking a wider... prep in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3...

  19. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...Advancement of Science, Rush Holt, followed by a networking reception. Saturday’s workshop will be an advocacy...laid out by a 2014 National Academy of Sciences working group (National Academy of Sciences. “The Postdoctoral... Advancement of Science, Rush Holt, PhD and a networking reception. Saturday’s workshop will progress ...

  20. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... me troubleshoot some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements...cells for infection If your transductions aren’t working, you can check if virus is being generated by your...template. Regardless of the type of virus you’re working with, there’s a lot that goes into virus design...

  21. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific...

  22. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...amount of virus you actually have in your prep. Taking time to properly titer your virus will not only...original plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will ...

  23. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...explain how it is interfering with your work. Avoid talking about your emotions, such as frustration or anger...

  25. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...in brackets and add the upstream and downstream flanking sequences to their input string (Figure 2). This...prediction models. In addition, we provide options for nicking gRNAs (used for PE3 and PE5 prime editing strategies...strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves time and resources by ...

  26. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...components of the indicator, which can make the kinetics of voltage sensing to signal output slightly slower...fluorescent signaler. The pro of this system is the kinetic response time from signal input to readable output...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and...

  27. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ... I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs...the viral particles of the AAV serotype you’re working with are primarily found in the culture media, .... If you’re new to creating gradients, practice making the iodixanol layers before doing your first purification...

  28. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ... localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by...fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery of many ...cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently tagged...

  29. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...the FP (fused to the protein of interest), thus tracking the localization of that biomolecule inside living...eel was shown to mature independently of oxygen, making suitble for use in anaerobic conditions. Maturation...fluorescent protein should I use?" And if you're looking for more fluorescence microscopy tools and techniques...

  31. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...our samples 1:6 million to 1:25 million.  After making the dilutions, they are transferred to a new plate...performing ddPCR Clean everything You must have a clean working area to titer your AAV by ddPCR. Droplet digital...generator, you should avoid pipetting too quickly when making your virus dilutions. This will reduce aerosols...

  32. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...domain proteins, they can be expressed in bacteria, making nanobodies a recombinant and renewable anti-IgG...available nanobodies from Pleiner et al. Before picking a secondary nanobody, it’s important to review ...expression plasmids here Additional Resources Looking for monoclonal primary antibodies to use with the...

  33. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...does not do). Furthermore, molecular biologists working with vectors are, in most cases, interested in ... the viral integrase protein (IN) during step 3 (King et al., 2008) (Hu & Temin, 1990). At step 3, the....1016/0022-2836(73)90277-5. PubMed PMID: 4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C....

  34. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...dye, mauvine, was discovered in 1856 by William Perkin. His discovery set off a flurry of activity and...labeling strategies and improved fluorophores are making chemical dyes increasingly attractive and accessible...Medical Institute. He would rather be in the lab making new fluorophores for advanced imaging modalities...

  35. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...speech, coordination, purposeful use of hands, and walking. They may exhibit slow head growth, develop seizures...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge O, Zoghbi HY (2013) An AT-Hook...

  36. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...zebrafish embryo develops outside of the mother, making them ideal for studying vertebrate embryogenesis... target mRNA inhibit its translation either by blocking initiation or splicing. (RNAi, in contrast, engages...unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow” fish lines have been...

  37. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...reliable and robust high quality viral vector service, making an array of ready-to-use viral vectors available.... I would be remiss to talk about them without looking at what they’re delivering and where they’re delivering...of our most popular ready-to-use viral vectors, making up almost 10% of our requests by 2019 (Figure 5A...

  38. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ... replication and regulation in bacteria Before looking at an example plasmid, let's review how E. coli...different ways: methylating newly synthesized DNA, blocking binding sites, or trading out ADP for ATP to re-activate...mRNA and prevent this pseudoknot from folding, blocking replication. Created with BioRender.com. Plasmids...

  39. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...information to explain why you are interested in working for that organization or group. This is particularly... of what to put in your cover letter, start by looking at the requirements in the job description. For... good place to state why you are interested in working for this lab and/or company. Remember to proofread...

  40. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ... delivery to only a few cell types.  If you’re looking for more control in the cell types your lentiviral...retrograde direction from the synapse to the cell body, making it a great tool to study neuronal connectivity....HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents...

  41. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...when 2 mismatches are present. The protospacer flanking sequence (PFS) for LshCas13a, which is analogous...have utilized dCas13 fusion proteins for imaging, tracking, modulating splicing, and regulating expression... in length and does not require a specific PFS, making it a very flexible cleavage system. LwaCas13a is...

  42. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ... of how they actually “report” as biosensors. Making aptamers glow The flexibility of aptamers means...fluorogen and increase its fluorescence, essentially mimicking GFP.  Though the aptamer, called Spinach, required... Eric J. Perkins...

  43. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...BP Reaction takes place between the attB sites flanking the insert and the attP sites of the donor vector...gene! STEP 2: Generate an Expression Clone When making the expression clone, it is important to choose...fragments, cDNA or Genomic DNA and is available for all kind of organisms from mammals to E. coli. Glossary of...

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

    Type
    Blog Post
    ... month without a new, drug-resistant ‘superbug’ making headlines in one part of the world or another. ...that only infect bacterial cells), or a bacterium picking up resistance genes from it’s environment (from...resistant bacteria to over-take the population and making them more difficult to treat in the future. Patients...

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

    Type
    Blog Post
    ...through smart engineering Different FLP versions – making FLP efficient The first version of FLP discovered...proteins needed for later experiments (cellular tracking for instance) can be controlled with FLP-FRT. ... Cardin, Jessica A. et al. 2009. “Driving Fast-Spiking Cells Induces Gamma Rhythm and Controls Sensory...

  46. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...a bit of tuning before you know your system is working as expected. Finally, for each gene you want to... scorer 1.0 (Now 2.0) (Chari et al., 2015) and picking whichever guides are closest to the transcription...Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, van der Meer D, Dow D, Buser-Doepner C, Bertotti...

  47. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...that particular region. No matter where you are looking, you can either scan the genome yourself for potential...leading to degradation that can set you back days. Working with plasmid DNA is much easier and doesn’t require...slightly different method to genotype their pups. Taking a toe, a finger, a tail tip, or a combination of...

  48. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...we are interested in circuits that mediate drug taking behavior. If a circuit reinforces behavior (activation...sense of pleasure that the animal perceives through taking a drug. The ideal behaviors to test reinforcement...amygdala to nucleus accumbens facilitates reward seeking." Nature 475.7356 (2011): 377-380. Pubmed PMID:...

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

    Type
    Blog Post
    ... this, similar systems have been developed for tracking various cell types and organisms and for improving...recombinase. The XFPs in Flybow are membrane-tethered, making this system useful for visualization of cellular...construct was then used to develop a system for tracking the fate of Langerhans cells. In the Ubow mouse...

  50. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...dependency on magnesium and higher thermal stability, making it a potentially more useful tool for in vivo studies...much potential for growth as RNA biology, we are looking forward to reports on the next additions to aptamer...bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology 9.10 (2014): 2412-2420....
Showing: 551 - 600 of 929 results