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We narrowed to 957 results for: REP

Showing: 541 - 560 of 957 results

  1. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    ...region. Thankfully, due to their prominence, the preparation of the fiber optics is probably the least difficult...effect is also an important consideration when preparing the implant itself, which brings me to the second...same considerations apply to the polishing and preparation of the patch cable (although you may be able ...wirelessly-controlled optogenetic implants have been reported by several groups [4, 5, 6]. These have the obvious...

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

    Type
    Blog Post
    ...sigh. The problem is that many of these studies report correlations without clearly testing whether a ... when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites blocked by dCas9....for dCas9 to interfere with DNMT1 even through replication. The dCas9 also needs to be removeable, otherwise...Targeted DNA demethylation in vivo using dCas9–peptide repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol...

  3. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    Blog Post
    ...for genome editing, activating gene expression, repressing gene expression, and much more. But there’s one...meningitidis (Nme) (Pawluk et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and... viruses. For example, an Acr gene was used to replace genes in the difficult-to-engineer Sulfolobus islandicus...viral particles that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-...

  4. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...mild symptoms. This changed in January with the report of an outbreak of the virus in Brazil that was ...-ready format when the Zika virus outbreak was reported, and we felt that, given the critical need for...such a short timescale. Our article ultimately reported the development of a rapid design and prototyping...Find the Zika Sensor Constructs at Addgene As we report, we have embedded this next generation of toehold...

  5. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...transfection). This differs from typical insect cell preparations, which we talk about below. Processing The viral...eliminate any HEK-293T cells. If you titrate your viral preps, lentiviral titration requires an additional couple...to collect your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in...Lentivirus Production Tips for Titering Your Lentivirus Preps More resources on Addgene.org Addgene's virus protocols...

  6. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ...annealing and extension is repeated 25-35 times to exponentially replicate the target DNA of interest....concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify DNA sequences ...Quantify RNA levels Molecular Diagnostics NGS library prep QC ddPCR Determine ultrasensitive and absolute...

  7. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's...FP dimerization. Equally problematic, several reportedly monomeric FPs are not actually monomeric, at ...Incorporation into a FRET biosensor A FRET biosensor is a reporter that undergoes a conformational change to bring...help you make better fluorescent fusion protein reporters for your studies and greatly lower your chances...

  8. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...bacterial DNA repair mechanism. The pathway is ubiquitous, with successful recombination reported in all laboratory...use of this pathway as a cloning tool was first reported over 25 years ago (Douglas and Howard, 1991, Bubeck...exploiting bacterial in vivo assembly." Scientific reports 6 (2016): 27459. PubMed PMID: 27264908. PubMed ...

  9. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...wild-type viral particle — and of the normal, replicative life cycle — can help you understand these seemingly...and viral vectors. Key to the wild-type virus’ reproductive cycle is the fact that the virus must have its...inside of the capsid . This is useful for viral reproduction, and for causing contagious infection; however...Hutcheson for her assistance with lentiviral replication.  Resources and References More resources on...

  10. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...he joined an elite club, becoming just the fifth repeat Nobel Prize winner in history. Interestingly, he...  Fig. 1: Undergraduates in Provost’s CURE prepare to analyze their recently purified MDH to study...results with the broader scientific community represents the culmination of many such experiences. For....     Fig. 2: Students in an MDH CURE replaced select residues of human mitochondrial MDH (hMDH2...

  11. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...up colony PCR reactions is nearly identical to preparing a standard PCR reaction: combine template, primers...other colony PCR tips are highlighted below. Preparing template: Pick a single colony with a sterile ... cultures with the clones you pick and only mini-prep the positive ones. Regardless of which method you... a few positive clones, the last step is to mini-prep these clones and submit the plasmids for Sanger ...

  12. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ... are institutions doing? Institutions are now preparing to comply with the changes to the law on Dec 1st...National Postdoctoral Association Institutional Policy Report 2014). However, ~30% set their minima lower. Furthermore... 340 institutions listed as having postdocs, representing ~85% of the postdoctoral workforce. Based on... the 2015 and 2016 Future of Research Symposia Prepare Yourself for a New Career in Science with Our Career...

  13. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...share- as opposed to sell- science. 4. You can prepare answers beforehand. Joanne answered many questions...questions to provide moral support and to monitor user replies, but I can’t thank Joanne enough for all the work...incentivize good mentorship: AMA mechanics: Be prepared! The mechanics of doing the AMA were interesting...interesting. I started watching the questions and preparing answers early in the day. Although I didn’t start...

  14. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...activity, such as Fos, Egr3, and Arc. Thus, CRISPRa represents a powerful method to dissect the individual and...astrocytes, and oligodendrocytes, provided that reporter genes specific to these cell types are used. The...including studying mRNA dosing effects, cellular reprogramming, and disease modeling. Further, the paucity ...identify factors that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed...

  15. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...expression using other methods.  In terms of preparation, VSV-G is stable and can withstand ultracentrifugation...The glycoprotein will be incorporated into the replicating rabies virus envelope, travel retrogradely, and...Lentivirus Get tips for titering your lentiviral preps Resources on Addgene.org: Check out the lentivirus...lentivirus guide Find ready-to-use lentiviral preps at Addgene ...

  16. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...), and width (W). The different measures are represented as FSC-A/SSC-A, FSC-H/SSC-H, or FSC-W/SSC-W, ...describe the shape of a histogram, which graphically represents the time and intensity of the cell’s illumination...cytometer (before you set the first gate) will be represented through FSC-A vs. SSC-A, so most people use that...scatter (FSC/SSC), the detected photons can be represented through a graph of photocurrent vs. time. The...

  17. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...Bunch of baby spinach, or BoBS, contains tandem repeats of Baby Spinach. It was developed in a study of...growth as RNA biology, we are looking forward to reports on the next additions to aptamer soup! References...fluorescent rna aptamer spinach2 for imaging of replication and immune-mediated changes in intracellular ...Spinach2 reveals the dynamic nature of trinucleotide repeat–containing RNA." Nature methods 10.12 (2013): 1219...

  18. Tips for Getting a Faculty Position

    Type
    Blog Post
    ...finer points of your techniques. You DO need to be prepared to know the literature of your field and all of...absolute minimum, can appreciate their research. Be prepared to discuss other people's research. Nothing turns...by email or in person. If you feel compelled to repay me in some way, I encourage you to help your friends...with their job searches. If you feel the urge to repay me personally, I will gratefully accept a very hoppy...

  19. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...often used for animal germline modification, to repair or add in disease-causing mutations, but, until...suited for genome editing. The homology-directed repair (HDR) pathway is downregulated in mature tissues...Although the three DMD studies referenced here represent a great step forward for CRISPR gene therapy, ... and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit Genes Read How Non-Homologous End...

  20. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...addition, PVDF membranes can be repeatedly stripped of an antibody and reprobed with different antibodies allowing... (SDS-PAGE) is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content...control. In these cases, the western should be repeated.  Finally, while westerns are a useful way to ...
Showing: 541 - 560 of 957 results