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We narrowed to 876 results for: tra

Showing: 461 - 480 of 876 results

  1. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...best gRNA Figure 1: Validation of double strand break (DSB) mediated homology dependent repair ...repair (HR, homologous recombination; SSA, single strand annealing) took place and reconstituted the EGFP...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...fluorescence, the better that gRNA is at causing a double stranded break at the target site, allowing the two halves...plasmid DNA is much easier and doesn’t require the extra precautions needed when dealing with RNA (e.g. face...(2014): 132-141. PubMed PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. ...

  2. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...have since been retracted due to cell line contamination: Epithelial Mesenchymal Transition Is Required for...records allows you to track a cell line’s age and monitor any changes. Your tracking system could be as ...biochemical assays followed by in vivo testing. The mantra being “you can’t trust those whacky cell lines”...colleague. Time and money are wasted, papers are retracted and there is a loss of reputation that can impede... good recordkeeping, you need to have a way of tracking how many passages the cells have undergone and...variety of pharmaceutical companies including GSK, AstraZeneca-MedImmune, and Infors & Applikon. Due to his ...his high level of experience he now travels around Europe giving talks and seminars on cell culture and ...

  3. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...has been removed/mutated to neutralize the nuclease activity. For strains that still carry this gene, ...from the DNA. These apurinic sites if in double stranded DNA are repaired by base excision repair, but ...but can lead to mutations in single stranded DNA undergoing replication.   DNase on the other hand can digest...main causes of degradation during plasmid DNA extraction and purification from E. coli is the presence...the gene endA. Nowadays, the most common E. coli strains used for plasmid amplification are endA negative..., there are usually extra steps like heat inactivation, protein degradation, and/or the use of chelating...dilute your sample in water to reduce the EDTA concentration. Alternatively, if the priority is to directly...

  4. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...cells, and someone new to plasmids may need some extra guidance to understand the specific components that...survival, such as an antibiotic resistance gene. In contrast, plasmids utilized in the lab are usually artificial...plasmids to self-replicate within a cell make them attractive tools for the life scientist or bioengineer. ...for further study. Promoter Region Drives transcription of the target gene. Vital component for expression...cell types. Primer Binding Site A short single-stranded DNA sequence used as an initiation point for PCR...vector containing the newly inserted gene is transformed into bacterial cells and selectively grown on...Produce proteins that glow so that scientists can track their location or quantity inside a cell Plasmids...

  5. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...fun is building within the constraints given. And there were many constraints during the COVID-19 pandemic...looking, I found an inexpensive HDMI transmitter that could transmit a signal between a wall. There was ...impressive, but due to costs and availability, a traditional solution was completely out of reach for Addgene... things first was getting that wireless HDMI transmitter/receiver, to beam the camera’s image to my remote...then set up a monitor connected to the wireless transmitter and an iPad in the office next door (connected...with wireless video receiver, camera control, and extra hand sanitizer.    Cycle 4: Venturing into the...was a bit cumbersome, but it was mobile enough to travel around the office with me, and to move it out of...

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

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...fluorescent protein that was derived from the tetrameric yellow fluorescent protein (LanYFP) from the ...fusion tag for a wide range of applications from traditional imaging to FRET and stochastic single-molecule...fluorescent protein mTPF1 is a monomeric version of the tetrameric cyan fluorescent protein from the coral Clavularia...Alberta and University of Oregon created mTFP1 by transforming E. coli with a gene library of approximately...approximately 484 variants, including some potential tetramer-breaking mutations. The library was then screened through...(2006): 531-540. PubMed PMID: 16859491. PubMed Central PMCID: PMC1698604. Ai, Hui-wang, et al. "Hue-shifted... 6.1 (2008): 13. PubMed PMID: 18325109. PubMed Central PMCID: PMC2292683. Shaner, Nathan C., et al. "A...

  7. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    Published
    May 6, 2014, 1:07 p.m.
    ...sensor proteins, particularly for the intracellular imaging of transition metal ions, such as zinc, copper,...field is a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors. Perhaps most ...firing in neurons and high resolution functional tracking of T lymphocytes, too, making them a versatile...developed and published in 2010 for visualizing the transcription factor cAMP-responsive element-binding protein...really important and when? That is not known.” Transition Metals Maarten Merkx of Technische Universiteit...the sensors involves plenty of trial and error. Transition metals also present challenges in that they are... are found in cells at much lower concentrations than calcium, requiring greater sensitivity. He and his...

  8. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    Published
    May 16, 2018, 1 p.m.
    ...FPbase Fluorescence Spectra Viewer can be used to compare excitation and emission spectra when available, ...soon feature filter spectra and collection efficiency calculations.  With ~80 FP spectra (and rapidly growing...than is currently represented at fpvis (complete spectral information, accession numbers, sequences and ...thinking about FPs, it would be great to have a centralized, community-editable resource for storage and ...the largest available online collections of FP spectra. Every protein has a page with all the information...biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools, and a ...

  9. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    Published
    June 18, 2019, 1:15 p.m.
    ... of the sensor. This “contrast” variant provides improved detection for tracking large populations of ...ppAAV-CKIIa-stGtACR1-FusionRed New tetracycline transactivators Plasmid Serotype Name 99120  AAV1...particular study, which includes comparing the transduction patterns of different AAV serotypes. To help... Biosensor AAV The ability to detect the neurotransmitters and other bioactive molecules is integral ...jGCaMP7c variant in AAV1. This variant exhibits high contrast between its resting state and its calcium-bound...fusion fluorescent proteins can affect DREADD trafficking and localization. The Sternson lab recently deposited...

  10. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ... antibody specificity. The gold standard in demonstrating primary antibody specificity is to test the ...and knockout samples are the gold standard to demonstrate primary antibody specificity. B, secondary antibody-only... secondary antibody specificity must also be demonstrated. Secondary antibodies have a tendency to interact...equal loading across samples and equal protein transfer from the gel to the membrane. A loading control...dead cells). Compensation controls address the spectral overlap that occurs during multiplexing. Compensation... Crossref, doi:10.1369/jhc.2010.956920. PubMed Central PMCID: PMC3201116. Hewitt, Stephen M., et al. “...Crossref, doi:10.1369/0022155414545224. PubMed Central PMCID: PMC4212362. Alexander, Steve P. H., et al...

  11. Clear and Accessible Writing

    Type
    Blog Post
    Published
    Feb. 15, 2024, 2:15 p.m.
    ...gradient tube before and after ultracentrifugation so that a scientist can extract virus from the correct layer...layer. The right panel illustrates a gradient tube before and after ultracentrifugation. The viral supernatant...from AAV Purification by Iodixanol Gradient Ultracentrifugation First, you’ll want to identify the reader...for AAV Purification by Iodixanol Gradient Ultracentrifugation. The reader may be, for example, a graduate... differently in a poster compared to an online abstract or Addgene blog post.  Here, we are presenting...profile photo of an iodixanol gradient after ultracentrifugation. The interfaces between layers are muddled...

  12. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    Published
    June 15, 2023, 1:15 p.m.
    ...factors can negatively impact both translation and fidelity of translation, leading to system-wide effects... incorporation of an unnatural amino acid in translation (Noren et al., 1989). In the late nineties, this...non-interacting with) the cells’ own natural translation machinery; these are often incorporated into ...complicated? Well, while the techniques are straightforward, the systems – the cells growing and producing...final protocol may involve a sensitive balance of trade-offs. This balance can be extremely tricky to find...for the phosphoserine and, because it uses ∆serB strains, does not require exogenously added phosphoserine...checking out all of the plasmids and bacterial strains (available at Addgene for GCE work. Plus, keep ...

  13. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    Published
    Aug. 19, 2025, 1:15 p.m.
    ...emission (solid) spectra for various teal, green, yellow, orange, and red fluorophores. The spectra for mOrange2...Simultaneous fluorescence (upper trace) and electrophysiological recordings (lower trace) using OCaMP expressed ...3: One- and two-photon excitation and emission spectra of OCaMP. 1P excitation peak is 545 nm, 1P emission...signal-to-noise ratio (SNR). Figure 4: Calcium titration curves for OCaMP and various red and yellow GECIs...expressed in mouse cortex. Bottom: Zoomed-in view of traces corresponding to the dashed box above. The number ...potential (AP) spiking events are indicated below the trace; asterisk indicates a single AP. B) Averaged fluorescence...

  14. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...how they started their company here. The first transgenic plant was engineered over 30 years ago, but plant... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...community needs to reclaim the term “GM” by demonstrating the current benefits and future potential of...decades, and the biochemical pathways involved in central anthocyanin metabolism are well understood. Additional...synthetic biology. Plant molecular biology has traditionally relied on overexpression and knockouts to better...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...

  15. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...used with a yeast strain deficient in that compound. Therefore known auxotrophic strain/ selection element...and animals. Unlike bacteria, yeast can post-translationally modify proteins yet they still share many of... genetic system, and a highly versatile DNA transformation system.Unlike most other microorganisms, yeast...plasmids oftentimes utilize an ARS to aid in high transformation efficiency; however, this region does not necessarily...resistant mutants and the insensitivity of yeast strains to some antibiotics. Auxotrophy is defined as the...compound required for its growth. Many auxotrophic strains of yeast exist which can be easily maintained when...antibiotic selection have made utilizing the more traditional drug selection methods feasible in yeast as a...

  16. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    Published
    Feb. 18, 2014, 4:22 p.m.
    ...group In my travels speaking to grad students and postdocs I have started to talk to trainees more about...better. Track your goal progress as a group. Practice your job talks for one another (trainees never get...read the book “Every Other Thursday: Stories and Strategies from Successful Women Scientists” by Ellen Daniell...experience. They make mentoring less mysterious and train the participants in skills useful for future mentoring... of people who are all interested in making a transition. Schedule your first meeting and get organized... your successes and support each other as you transition to the next stage in your careers! Stay in touch...

  17. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ... repeat region associates with the trans-activating crRNA (tracrRNA) which interacts with the Cas9 protein...experiments allows you to visualize transfection efficiency and even track localization in cellular experiments... the crRNA or the tracrRNA. Of course, in a single guide system (crRNA and tracrRNA as a single molecule... important cellular RNA molecules with post translational modifications. One of the most common of these...amide linkages, unlocked nucleic acids, and a constrained ethyl.  Can’t decide which RNA backbone modification...

  18. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    Published
    Oct. 1, 2019, 1:21 p.m.
    ...of using social networking to start, build, and track professional relationships. Here are my seven best... for example). This is also a great way to keep track of how you met the connection, as these notes are...trolls. People don’t like to connect with complete strangers as it borders on my first rule of networking--...a useful way to build or use a network. Total strangers can’t really help or refer you. Do not ask people... lot about requesting connections to complete strangers for informational interviews. Many have had success...considered intrusive and it’s still connecting with a stranger so is highly discouraged. You should be using ...my paper resume). It’s also a great way to keep track of past accomplishments. Follow all the rules of...

  19. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...wash steps often include low concentrations of imidazole. The concentration of imidazole used in the wash...Oftentimes, they rely on protein purification. In this strategy a single protein can be isolated from a complex...prevent it interfering with protein function. In contrast, the polyhistidine tag is very small and rarely...empirically determined. In addition, some metal ion substrates, such as those containing Co2+, have a lower ...affinity for polyhistidine than more commonly used substrates such as Ni2+ and are therefore more selective... systems, consider using these more selective substrates along with stringent wash conditions.   Find ...tightly the protein will bind to the metal ion substrate. If your protein of interest is expressed at a...

  20. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-linear ...important since different fluorescence imaging strategies use widely varying amounts of excitation power...levels (for a prolonged time). Other illumination strategies, such as 2-photon-excitation, selective plane...conditions such as cellular redox state and oxygen concentration may affect photobleaching rates (Shaner et al...the conclusions reached at high power may not translate to real applications in which far less power is...proteins. To avoid diffusion, the proteins are (i) trapped in microdroplets in a water/oil emulsion, (ii) ...) embedded in a gel, or (iii) attached to a substrate. These in vitro methods allow for a well-controlled...
Showing: 461 - 480 of 876 results