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We narrowed to 303 results for: hal.2

Showing: 201 - 240 of 303 results

  1. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic... Ononiwu, F., Pal, D., Manikas, J., Taveras, C., Hall, N. A., Silva, J. D., Freshour, J., & Hehnly, H....

  2. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach...antibody selectivity. Additionally, it can be a real challenge to troubleshoot RNAi failures. Potential explanations...cell survival or proliferation, an additional challenge is that only partial or ephemeral knockdown will...confirmation of antibody selectivity even more challenging. Correlation with RNA or Proteomic data from ...supportive of their specificity for the protein. A challenge to this approach is that the identity of the epitope...the specific sample and protocol. This can be a challenge to confirm. As such, the approach used may need...

  3. Hot Plasmids - February 2022

    Type
    Blog Post
    ...acceptable variations in DR sequences. Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes...SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag. ...screening technique, researchers identified a synthetic lethal interaction between MARCH5 and WSB2 genes using...fusion proteins, such as Cas9 and Cas12a, creates challenges for in vivo delivery and cell engineering applications..., is composed of only 529 amino acids, less than half the size of either Cas9 or Cas12a. Despite its small...

  4. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    ...perceived climate change risks. Nature Climate Change, 2(10), 732-735. doi:10.1038/nclimate1547 Kunda, Z. (...sources of information if those sources affirm or challenge their beliefs. I asked participants to read short...read an abstract for a study with findings that challenged their beliefs, they were much more likely to ...a phenomenon called “motivated reasoning,” the catchall term for a range of neat little tricks our minds...

  5. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...and trophies are awarded and coveted. I’ve only won 2 trophies over the years, which is either an indication...different labs to chat informally about their challenges, successes, and expertise. Who knows what could...

  6. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. 2013 Dec 2;8(12):e82009. doi: ...problem that he considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors...trial and error. Transition metals also present challenges in that they are found in cells at much lower...

  7. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...seqWell, Inc., our mission is to overcome crucial challenges in NGS by developing technologies that can help...problem in multiplex NGS library preparation. The challenge here is that, as more samples are sequenced simultaneously...

  8. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive...each half with PhyB or PIF6, phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In ...identical in sequence to the unmodified enzyme. Each half of the base editor is fused to a fast-splicing split...induces PhyBΔ and PIF6APB binding, bringing the two halves of Cre together. Image from Eisenhoffer et al.,...

  9. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...paper at Addgene. Other relevant blog posts 3 Challenges in Plant Synthetic Biology Quick, Versatile Plant...

  11. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ...still depolarize neurons at lower MOIs. Figure 2: ChRgers are delivered by intravenous injection into...optogenetics.  Challenges of engineering channelrhodopsins There are three main challenges to engineering...overcome channelrhodopsin design challenges To overcome these challenges, the Arnold and Gradinaru Labs ...lower multiplicity of infection (MOI) makes it challenging for ChR2(H134R) to function because it’s lower...

  13. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand...number of tandemly expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally...quantification of enhancer activity have remained challenging due to the variable availability of enhancer ...pTALYM3, pTALYM4 Boris Greber pTAL7a, pTAL7b Michal Zurovec pBlue-TAL Nathan Lawson pJDS Series ...degredation is the speed at which it works--typical half lives of AID-tagged proteins after the introduction...

  15. Bringing Sustainable Practices to the Lab: Recycling

    Type
    Blog Post
    ...the world’s population, we account for an estimated 2% of global plastic generation. Less than 9% of plastics...Labconscious. If you’re inspired by this post then I challenge you to take to action! These days, it’s important...

  17. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and...constructs, localization of the Rluc8 was more challenging. The Roth lab used structure-guided protein engineering...

  18. 10 Ways to Share Your Science!

    Type
    Blog Post
    ...but also something about how science really works. 2)   Read science writing in magazines or newspapers...   Consider participating in a future SciFund Challenge. While the goal is ostensibly to get some funding...

  19. RUBY-Red Siliques

    Type
    Blog Post
    ...make it easier to study these mutants.   Fig. 2: (A) Betacyanin is produced from tyrosine by the activity... positive plants, but screening these can be challenging and requires specialized equipment. Reporters...

  20. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ... temporal activation of a transgene. Figure 2: FLEX Cre-Lox system.  WPRE The Woodchuck Hepatitis...sequencing conditions which can read through this challenging sequence. Transgene The transgene is the gene...

  22. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ... sets Taking the time to identify and research 2 – 3 potential professions will highlight which skills.... No brainer! Engaging audiences from across the hall, fields, and other professions increases your chances...

  23. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges of succeeding in science...funding, bioethics in the age of CRISPR, and the challenges of science becoming increasingly politicized....ramifications. Readers unfamiliar with academia’s challenges with publication and funding may be dismayed ...The three chapters in this section describe the challenges of trusting science in a “fake news and post-...describes the author’s thoughts on science’s biggest challenges over the coming years. It is arguably the most...the book? It doesn’t need to. The problems and challenges described in The State of Science have not changed...may be the introduction they need to grasp the challenges you face in your career.  Acknowledgements: ...

  24. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes, ... making it all work – most of the time. #1 Thou shalt find enjoyment in your work If you are happy at ...and with attention to those around you #4 Thou shalt not bear the burden alone You, alone, are not solely...absolutely equally, which means you only have one-half of the guilt. You can handle that, right? Big tip...get enmeshed in a serious relationship. #5 Thou shalt not sweat the small stuff The house can be messy...yourself; you will make progress over time. #6 Thou shalt throw money at the problem If you can, pay someone...to relax and be with friends and family. #7 Thou shalt create community to help Involve family. If you ...

  25. CRISPR in the Clinic

    Type
    Blog Post
    ... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...Disease (SCD) and Transfusion-dependent beta thalassemia (TBT), both of which affect hemoglobin. Most ... enzymes target several strains of E. coli and lethally chop their genomes into many, many pieces. Locus...technology is rapidly evolving, and many of the challenges with therapies involve patient safety, effectiveness...

  28. Antibodies 101: Epitope Tags

    Type
    Blog Post
    ...is the most charged, with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been ...doesn’t come with a high background can be a huge challenge. Epitope tags eliminate the need for target-specific...

  29. Advice for Moving Into Sales After Your Science Postdoc

    Type
    Blog Post
    ... and whether I would be able to carry out a yeast-2-hybrid library screen; instead I thought they would...was researching, but I decided I needed a new challenge. Looking for sales and marketing jobs Like most...

  30. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ... remember that a resume is typically no more than 2 pages long. Write your resume for the job posting ... one, one of the biggest changes (and perhaps challenges) is the need to present yourself using a resume...

  31. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...research project. In total, the interviews lasted from half to all of the day.  This whole process may seem ...carefully. And even if you don’t have technical challenges, your interviewers might.  You’ll probably still...

  32. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...interferon gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of... of certain tumors. However, antibody stability, half-life, tumor penetration, inadequate concentration...

  33. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...establishing the dynamic range of your FRET assay (Figure 2). These labeling methods are freely interchangeable...PMID: 20161833. PubMed Central PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding...FRET distance measurements (for an example see Mahalingam et al., 2014).  Many thanks to our guest blogger...

  34. Hot Plasmids - November 2023

    Type
    Blog Post
    ... vivo and in vitro applications.   Figure 2: Modular features of DIRECTED particles where the ...externally-timed bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record...local proteome, but some applications have been challenging or inaccessible. The Ting lab recently made two.... B) HEK cell with an iPAK4 fiber labeled with HaloTag-iPAK4. Scale bar 10 µm. C) Images of iPAK4 fibers...fibers in cultured rat hippocampal neurons. HaloTag dyes are switched from yellow to red at 0 h and back...

  35. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...integrates into the trnS-leu2 locus.   Figure 2: Introducing Mobile-CRISPRi into bacteria. Top: A ...which creates a challenge when studying them since their complete knockdown is lethal. Incomplete or titratable... to lab-adapted bacteria because it has been challenging to introduce CRISPRi machinery into diverse bacteria...

  36. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...system (by measuring the fluorescence intensity); and 2) to visualize the localization of the FP (fused to...be split into two halves, which are non-fluorescent on their own. If the two halves are in close proximity...determine the proximity of two proteins fused to the halves of the split FP. This technique is also is also...

  37. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4... tool by splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein ...With the split system, only cells that have both halves of cGAL can drive expression of your gene of interest...

  38. Hot Plasmids - October 2020

    Type
    Blog Post
    ...products have distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different...synaptic connections and physiology is particularly challenging as they are great in number, generally very small...H2B-FT-red is long lasting and was shown to have a half life of 84 hours in primary mouse embryonic fibroblasts...

  39. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ...out for help so you can get the support you need. 2) Prepare an agenda for every meeting Supervisors are... of your supervisor’s management style, bring challenges to his or her attention as soon as you can. It...mutually beneficial solutions. If you package your challenges with your supervisor into learning opportunities...

  40. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first... Glucose Glucose, methanol, glycerol Heterothallic mating Efficient Efficient Inefficient...
Showing: 201 - 240 of 303 results