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We narrowed to 303 results for: hal.2

Showing: 141 - 160 of 303 results

  1. 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,...

  2. 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...

  3. 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...

  4. 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...

  5. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...

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

    Type
    Blog Post
    ...implies that reducing the power 2-fold does not reduce photobleaching by half. How exactly the photobleaching...prolonged time). Other illumination strategies, such as 2-photon-excitation, selective plane illumination, TIRF...described by the t½, which is the time after which half of the initial fluorescence intensity remains. The...

  7. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...TniQ for targeting (Halpin-Healy et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...proteins to form the Cascade targeting complex is a hallmark of Type I CRISPR-Cas systems, in contrast to single-protein...creates a 5-bp target-site duplication (TSD), a hallmark characteristic of the Tn7-family of transposons...follow him on twitter @Vopinator.     References: Halpin-Healy TS, Klompe SE, Sternberg SH, Fernández IS...doi.org/10.1038/s41586-019-1849-0 Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH (2019) Transposon-encoded...

  9. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip..., accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/Cas moieties carry out these functions...defense against invading phages. Rising to the challenge - Evolution of anti-CRISPR genes in phages The...

  10. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor Partnership...distribution after we partnered with them (Figure 2). In others, the distributor helped solve an issue...temperature for the duration of their journey. Figure 2: Annual distribution of items to countries with a ...persisted despite our best efforts. With a little over half of our orders coming from researchers outside the...

  11. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126... Zhong, B. L., Elliot, J. M., Wang, P., Li, H., Hall, R. N., Wang, B., Prakash, M., & Dunn, A. R. (2024...

  12. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi....WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang...

  13. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... than control, 3; same as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice...stranded break at the target site, allowing the two halves of EGFP to recombine by homology directed repair...

  14. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...small operation, so sometimes it can be a big challenge. But ask any filmmaker or videographer and they...

  15. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ... fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil... lymph nodes at different times after antigen challenge, scientists were able to track where GFP labelled...et al. 2016, Cavanagh et al. 2008)? Technical challenges of two photon microscopy Intravital imaging is... at steady state or under physio-pathological challenge. However technical specificities have to be taken...

  16. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...the University of California Berkeley. It’s been half a year since the march for science on April 22, ...present at the lab bench," Liu, a UM undergraduate, challenged us. I worried the march would be a simple, unquestioning... fingers of skepticism to defend an agenda that halts current data collection, destroys existing data,...

  17. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation signal directs...processing and contributes to variability in RNA half-life, and ultimately gene-expression. Polyadenylation...39322032   No terminator is 100% efficient at halting transcription of the template and initiating the...

  18. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...FRET signal back again. The Ting lab constructed 2 versions of this reporter to target the K9 (pcDNA3... 525.7570 (2015): 533-537. PubMed PMID: 26352475. 2. Bygren, Lars O., et al. "Change in paternal grandmothers...real-time, posing a huge challenge to studying epigenetic changes over time.  This challenge was difficult to ...

  19. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ... environmental sustainability in mind (Table 2). Table 2: Sustainability features highlighted in plasmid...debris”; and then clean-up the plasmid DNA (Figure 2). For most kits, the steps for lysing cells and removing...together — no mixing and matching!   Figure 2: Plasmid isolation workflows are similar across kits...inks Reduced cardboard packaging by more than half 70% recycled materials Sustainable forestry ...

  20. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...guide sgRNA-Sirius-8XMS2 which will recruit the HaloTag with red fluorescent ligands to the loci of interest...
Showing: 141 - 160 of 303 results