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We narrowed to 297 results for: hal.2

Showing: 121 - 140 of 297 results

  1. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...columns are shown in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first... (budding yeast), S. pombe (fission yeast), A. thaliana (mustard weed), O. cuniculus (rabbit), synthetic...

  3. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ... characteristics of select AAV serotypes in Table 2 from the paper. Delivery of AAV into the brain can...routes for neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific...development, learning, and behavior poses many challenges in neuroscience with many brain regions, neuronal...

  4. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... 10 90 2 x 10 8 10 of 2 x 10 8 dilution 90 2 x 10 7 10 of 2 x 10 7 dilution 90 2 x 10 6 10 of 2 x 10 6...6 dilution 90 2 x 10 5 10 of 2 x 10 5 dilution 90 2 x 10 4 10 of 2 x 10 4 dilution 90 2 x 10 3 Pro-Tip...molecules/μL To obtain a solution at 2 x 10 9 molecules/μL: 1.59 x 10 11 / 2 x 10 9 = 79.8X dilution ...your standard curve plasmid (2 x 10 9 stock made in step #1): Volume of 2 x 10 9 stock or previous dilution...stock 45 uL 10X 10X Dilution 2 5 uL Dil. 1 95 uL 20X 200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution ...time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment ...single channel pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...

  5. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...required components are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation...gone through an additional round of evolution (AID-2) to reduce some of the leakiness associated with the...Nabet et al., Chemical Biology, 2018.    HaloTag The HaloTag was initially developed as an affinity tag...the Halo-tagged protein in question, it is targeted to the proteasome for degradation. The HaloTag is ...straightforward system – HaloTag and its PROTAC are all that is needed to facilitate degradation. HaloTag is also the...visualization of proteins of interest. A fluorescent Halo ligand can be added to media, buffer, etc., which...proteolysis-targeting chimera (PROTAC) was developed for HaloTag. PROTACs are engineered molecules which target ...

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

    Type
    Blog Post
    ...is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation...since the Future of Research Symposia Ongoing challenges facing the scientific enterprise and the way ...landscape of the scientific enterprise will provide challenges and opportunities for us all.  For example, implementation...categorize all postdocs at institutions, will be challenging. At the same time, there is much opportunity ...the pressure on for further increases will be challenging, but will be aided by the development and implementation... is an important way to push the balance from challenge to opportunity. We hope to see you at the upcoming...

  7. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    ...groups, such as their vaccine production, SARS-CoV-2 research, or adenovirus packaging cell lines dataset...-Line Knowledge Resource. J Biomol Tech. 2018; 29(2): 25-38....split you got from the graduate student down the hall who got it from some technician downstairs who got...

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

    Type
    Blog Post
    ...base pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles that...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-Cas system. 5. ...

  9. Antibodies 101: Flow Compensation

    Type
    Blog Post
    .... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations.   Figure 2: Compensation example based on experimental sample..., you will see the populations move, as in Figure 2, following compensation. Figure 3: Flow plot...Compensating between your fluorescent markers can be challenging! But we hope this guide can help you get started...

  10. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup... the body or self antigens, and toxins that are lethal to animals.  In the clinic, scFvs’ size also provides...plasmid in bacteria (Ahmed et al., 2012). One challenge of bacterial expression is the proper formation...

  11. CRISPR Guide

    Type
    Guide
    ...left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only...systems enable researchers to target anywhere from 2–7 genetic loci by cloning multiple gRNAs into a single...Cas9 is included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with...the presence of infectious organisms (like SARS-CoV-2 ) and genetic mutations. Similar to Cas9 and Cas12...Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...off-target effects by using a single Cas9 nickase and 2 different gRNAs, which bind in close proximity on ...

  12. Lentiviral Vector Guide

    Type
    Guide
    ...vectors. Viruses , 3 (2), 132–159. https://doi.org/10.3390/v3020132 PMID: 22049307 Ghaleh, H. E. G., Bolandian... Therapy , 24 (2), 132–142. https://doi.org/10.1089/hum.2012.229 PMID: 23311447 Shalem, O., Sanjana, N...intact. First-generation plasmids include (Figure 2): Transfer plasmid — contains transgene and wild-type...envelope removed Envelope plasmid — contains env Figure 2: First-generation lentiviral plasmids. Created with...plasmid 1 — contains gag and pol Packaging plasmid 2 — contains rev Envelope plasmid — contains env Third-generation...Infectivity and Integration Targeting. PLoS ONE , 2 (12), e1340. https://doi.org/10.1371/journal.pone.0001340...Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production Your Lentiviral Plasmid...

  13. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...PMID: 24086725. PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher...makes generation of high titer Cas9 lentiviruses challenging. This cytotoxicity also causes growth rate variability...):337-47. PubMed PMID: 12595892. 4. Sanjana NE, Shalem O, Zhang F. Improved vectors and genome-wide libraries...

  14. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET... developed in human cells, has been uniquely challenging to apply in dicotyledonous plants, or dicots....

  15. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires a less involved setup when working...infection rates in your experiment. Figure 2: Visual representation of common AAV serotypes used..., 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández, A., Low, S. H., François, A.,.... V., Teyssier, N., Pan, A., Danner-Bocks, S., Sanghal, T., Yadanar, L., Tian, R., Ma, K., Palop, J. J...

  16. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https...Come From and What Do They Mean? Overcoming the Challenges of Lentivirus Production Tips for Titering Your...

  17. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Arabidopsis thaliana seedlings following floral dip transformation’, Plant Methods. BioMed Central, 2(1), p....PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ...p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed Central PMCID: PMC1636043. 3. Kyndt,...

  18. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ... and what can be left out, which can be a big challenge!  The resolution Throughout your conflict, you...

  19. AAV Production in HEK293 Cells

    Type
    Protocol
    ...430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 Heat-inactivated...: 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following to the 2 L sterile bottle: 1836 mL deionized... solution can be stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working... mL of PBS. Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding...Cell-Stack (CS5) (Link opens in a new window) (3,180 cm 2 - the same surface area as 21 x T-175 flasks). Cell...flasks. Workflow Timeline Day 0: Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...

  20. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...without leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing...with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D...CRISPR/Cas9, TALENs, and ZFNs). However, several challenges still remain, such as making gene edits completely...nuclease technology with the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon...
Showing: 121 - 140 of 297 results