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We narrowed to 313 results for: POR C

Showing: 101 - 120 of 313 results

  1. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...store them in a -80°C freezer to prevent RNA degradation. They will degrade in a -20°C freezer.   RNA extraction...DEPC. Alternatively, glassware can be baked at 180°C for at least 4 hours.   If your final RNA sample(s...extractions- if you need really clean RNA, it’s important to make sure that when extracting, to only take...

  2. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...cloning method with high throughput capability. Engler C, Kandzia R, Marillonnet S. PLoS One 2008;3(11):e3647..., Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic...2011 Apr 14. PubMed.   Golden Gate cloning. Engler C, Marillonnet S. Methods Mol Biol. 2014;1116:119-31...like Gateway cloning or Gibson assembly. Another important consideration is the design of flanking overhangs...

  3. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...fragments: N-terminal BirA* (N-BirA*) and C-terminal BiraA* (C-BirA*). These two fragments are fused to...also which fragment of BirA*, either the N- or the C-terminus, the bait proteins were tagged with. It’s...interacting proteins. No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “... lysate fractionation. Important considerations for Split-BioID It’s important to remember that with Split-BioID...this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent...The Split-BioID method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol... other PPI methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins...

  5. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...length of your assay. The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most...to incubate their primary antibody overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature...the DIY method. As some proteins aggregate at 95 °C, you may want to check for any available data on the...expensive, reduce protocol time, can be stored at 4 °C, and are very consistent gel-to-gel.  Ladder Whatever...stored at your bench during the procedure and at 4 °C when you’re not running a western. They are not very...will noticeably degrade after several months at 4 °C.  Pro tip! If you get a speckled image, your protein-based...variety of pore sizes. If your protein of interest is small, you may want to consider a smaller pore size (...

  6. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...Williams, Eric J. Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado,...discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific...Hybridization (WISH) WISH is a technique that detects the temporal and spatial localization of mRNA transcripts in...

  7. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...still unexpectedly affect nearby genes (Figure 1A-C). Find CRISPR plasmids at Addgene! Knock-in: Introducing...site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (... doi:10.1080/15476286.2014.996067 Basak J, Nithin C. Targeting Non-Coding RNAs in Plants with the CRISPR-Cas...experiments are important for visualizing ncRNA localization and function, but directly incorporating very long...transcribed but do not encode proteins. They play important roles in many cellular and regulatory processes...complex and not yet fully understood, so it is important to carefully select the target locations of knock-in...nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription machinery of a target...

  8. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...   References and Resources   References Beck C, Zhang D, Gong Y. Enhanced genetically encoded voltage...: 10.1016/j.cobme.2019.10.010. Bando, Y., Grimm, C., Cornejo, V.H. et al. Genetic voltage indicators.... 71 (2019). doi:10.1186/s12915-019-0682-0. Beck, C., Gong, Y. A high-speed, bright, red fluorescent voltage...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...rhodopsin proteins were initially evolved for ion transport, not fluorescence, they are not nearly as bright...for penetrating deeper tissues depths (1 mm), important for certain applications such as in vivo tissue...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from ...

  9. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...doi.org/10.1038/mt.2009.280 Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski....2010.205 Trapani I, Colella P, Sommella A, Iodice C, Cesi G, Simone S, Marrocco E, Rossi S, Giunti M, ...Trapani I, Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella A, Colecchi L, Rossi S, Simonelli...1964 . https://doi.org/10.1172/jci34316 Bak RO, Porteus MH (2017) CRISPR-Mediated Integration of Large ...Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756 . https://doi.org/10.1016/j.celrep.2017.06.064...

  10. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...combs. To make genetic crosses properly, it is important to isolate females from males while they are still... after hatching into their adult phase. It is important to clear out the parent crosses from the container...feature when setting up genetic crosses. More importantly, this knowledge will stop you from embarking ...

  11. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...Adenine base editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464...Lee S-J, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G (2020...selection of guideRNAs. Of particular note, ABE8s reported in this work did not cause genome-wide, guide-...guide-independent off-target deamination, an important attribute for therapeutic application. ABE8s demonstrate...

  12. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce a differentiated somatic cell to...additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming ...22645348. PubMed Central PMCID: PMC3390875. 25. Patsch, C., et al., Generation of vascular endothelial and smooth... PubMed Central PMCID: PMC4363833. 35. Eguizabal, C., et al., Dedifferentiation, transdifferentiation,...1): p. 82-94. PubMed PMID: 23329641. 36. Jopling, C., S. Boue, and J.C. Izpisua Belmonte, Dedifferentiation... adenovirus [5], Sendai virus [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]...insulin-producing β-like-cells [37, 38]. In 1987, Davis et al. reported one of the earliest examples of transdifferentiation...

  13. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ... should already be at 37 °C. Most labs have a water bath constantly at 37°C for this purpose.Learn about... Cells usually have their comfort zone around 37 °C and 5% CO2 and get agitated with sudden changes in...passage are too young and should not be used for important transfection experiments right after they’ve been...

  14. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ... P., Rodriguez-Aguayo, C., Walton, B. L., Sood, A. K., Greaver, J., Janssen, C. F., & Lopez-Berestein,... Academic Press. Turner, P. V., Brabb, T., Pekow, C., & Vasbinder, M. A. (2011). Administration of substances...muscle groups (usually the thigh), though it is important to avoid injection into the sciatic nerve. Mice...

  15. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...assembly of NgAgo/ssDNA requires incubation at 55 °C - a dangerous, non-physiological temperature for mammalian...genome or just after a stop codon (as required for C-terminus epitope tagging). NgAgo survey Since it was..., he tries random things (101 lessons on Spanish, C++, guitar, photography) and often dreams of a sunny...Researchers from a variety of institutions recently reported their inability to recapitulate the results of...

  16. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...GFP (sfGFP) and mNeonGFP can fold in <10min at 37°C, mCherry takes ~15min, TagRFP ~100min and DsRed ~10hours...instance, enhanced GFP (EGFP) was optimized for 37°C, and is therefore most suited for mammalian or bacteria...GFPS65T is better suited for yeast studies (24-30°C). Brightness: Brightness is a measure of how bright...temperature). pH Stability: This parameter is important if you are planning to express the FP in acidic...

  17. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled proteins identifies proteins...bioRxiv (A) and Qin and Cheah et al. 2023 bioRxiv (B-C). They were able to label proteins moving to and...fiber labeled with HaloTag-iPAK4. Scale bar 10 µm. C) Images of iPAK4 fibers in cultured rat hippocampal... editing efficiencies in vitro and in vivo. Incorporating reverse transcriptase (RT) variants, PE6a and...., 2023, reused under CC-BY 4.0 License. Importantly, even commercial antibodies can be tethered on... more. For added precision, the authors also incorporated LOV-Turbo into TransitID experiments.  Find ...in light-regulated LOV-Turbo enables precise spatiotemporal control of proximity labeling in living cells...

  18. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

    Type
    Blog Post
    Published
    June 8, 2023, 1:15 p.m.
    ...References and Resources References Radtke, A. J., Chu, C. J., Yaniv, Z., Yao, L., Marr, J., Beuschel, R. T.... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv...along with the ability for any user to upload supporting documentation that agrees or disagrees with the...

  19. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...site for a total of 2 binding sites per antibody. C) IgG’s are big enough that multiple labels (green ... IgG secondary antibody (Pleiner et al figures 4A-C). And since they have limited cross-reactivity between...Bates, M., Böhning, M.I., Chug, H., Görlich, D., Lee, C., Pleiner, T., Schliep, J.E., Stark, H., Trakhanov... genetic engineering of a tag or fusion with a reporter enzyme. For multicolor staining, careful consideration...al. Before picking a secondary nanobody, it’s important to review a few characteristics of your primary...

  20. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... Cas9 into n-terminal (Cas9(N)-2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA ...obtain even more precise temporal control of gene knockout or activation, the c-terminal cas9 fragment ... the nuclear export sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS ... (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(N)-FRB-NES). This system provides users greater temporal control ...an n-terminal DNA recognition domain and a mostly c-terminal nuclease domain. Feng Zhang's group utilized...fragment was fused with FK 506 binding protein 12 (Cas9(C)-FKBP-2xNLS) and the n-terminal cas9 fragment with ...constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab of Tomás Santalucía...
Showing: 101 - 120 of 313 results