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We narrowed to 367 results for: proc.2

Showing: 101 - 120 of 367 results

  1. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    Published
    Feb. 11, 2014, 12:13 p.m.
    ...require application submission in-house approximately 2 weeks before the RFA due date in order for the administrative...deliverables to the aims requested in the call. Step 2: Develop a research plan Most grant applications ...proposal typically lists the deliverables, explains 1-2 potential pitfalls of the intended protocol, and offers...want to mentor you during the grant application process. When my PhD advisor first asked me to write a ...PI asks for assistance in the grant application process, she has faith in you! Additionally, keep in mind...motivating research will help you understand the process and continue to improve your grant writing skills...Still want more information on the application process? What types of writing feedback do you find most...

  2. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ... fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly to...PMID: 24194740. PubMed Central PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein...PubMed PMID: 10500161. PubMed Central PMCID: PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators...orientation of the FRET pair. Since many biological processes occur within the typical FRET range, FRET efficiency...fluorophore on the same protein and are useful when a process affects the conformation of the probe. More on ...answer sophisticated questions about cellular processes. Currently, FRET experiments can probe protein-protein... activity of small molecules, detect cellular processes and signaling cascades, quantify mechanical tension...

  3. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...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...guide to our online data entry process. Figure 1: Begin the deposit process by clicking “Submit Online Deposit... our deposit spreadsheet?  Our plasmid deposit process can be broken down into four simple steps: Send... contact you if any issues arise during the QC process. As the depositing scientist, you will have the...information in a deposit spreadsheet. The data entry process On the Addgene home page, if you scroll over “Deposit...

  4. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...purify IVT sgRNA in the lab. Figure 2: Cas9 RNP complexed with 2-part cr:tracrRNA tags genes more effectively...into stem cells. Cells are allowed to recover for 2-5 days and then analyzed for tagging efficiency using...IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient...cell suspension, transfect and let cells recover for 2-5 days.Note: We use the Nucleofection method for delivery...success was ~%50. If you get low knock-in efficiency (<2% positive by ICC for the tag) or do not see any cells...PMID: 28096221. PubMed Central PMCID: PMC5312033. 2. Dewari PS, Southgate B, Mccarten K, Monogarov G, ... Encouraged by the high efficiency and easy procurement of cr:tracrRNA reagents, we scaled up these tagging...

  5. Year of the Camelid: Antibody Style

    Type
    Blog Post
    Published
    May 9, 2024, 1:15 p.m.
    ...and a source of sdAbs (Figure 2) (Xu, et al., 2021).      Figure 2: Genetically modified mice offer... from camelid mice and llamas neutralize SARS-CoV-2 variants. Nature 595:278–282. https://doi.org/10.1038...? In the early years of sdAbs development, the process started with immunizing a camelid (usually llamas...

  6. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...detect your targeting vector or gene of interest. 2. Breed your parental mice to generate F1 Once you’...maintain your parental strains throughout this process. In the F1 and later generations, mice that are...

  7. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ... Opto-Cre-Vvd is fast and can cut in as little as 2 hours.  Sheets, MB, et al., ACS Synth Biol. 2020....they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several reasons...a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where B is G, T, or...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a ...active site. The RuvC active site in Casɸ can both process crRNA and carry out the crRNA-guided DNA cutting...

  8. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    Published
    Oct. 22, 2014, 1:57 p.m.
    ... let the panelists introduce themselves. I do a 1-2 minute introduction. If you chose good panelists, ...take 5 minutes to introduce themselves and, voila, 1/2 of your time is gone with no interesting discussion...introductions. The moderator asks questions, calling on 1-2 panelist for each question in a way that elicits those...shift, they are checking out. If you can, plant 1-2 good questions with trusted audience members. A good... Your audience may have a lot of real or self-proclaimed experts on the topic at hand. You can use this...

  9. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ...are many docking sites available for integration. 2. How soon would you like your flies ready for use?...beautiful immunofluorescent images like in this paper (2). Metabolic assays such as Seahorse can be employed...PMID: 16547094. PubMed Central PMCID: PMC1526508. 2. H. Jiang,  et al. Cytokine/Jak/Stat signaling mediates...make a transgenic fly in order to study it. The process is fairly straightforward. Basically, you place... incorporated into the germline. To begin this process, you will first need to do some molecular cloning...

  10. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...extracted the DNA and processed the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly... potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual...quality, we undertake a rigorous quality control process. In 2016, we launched our viral service to continue...bacteria snuck into the prep during the production process? These questions are especially important when ...CHRM4 gene. When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing...answer your questions about our quality control process! The images in this blog post were created using...

  11. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ... components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or module...overhang specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual... (KO, CRISPRa, CRISPRi, editing)   Figure 2: Selecting module fragments on the Fragmid website...Fragmid. Fragmid is a modular vector assembly process designed for a quick, plug-and-play approach to...of the design work out of the plasmid creation process. Each module of Fragmid represents a class of components..., the Fragmid website, which breaks the design process down into four easy steps, provides the real value...

  12. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating... characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver... its expression. More specifically, during the process of RNAi, an enzyme dubbed Dicer cuts or “dices”...Phagocata morgani and Phagocata gracillis, the process is a little more complicated. These strains are...RNA inhibits gene expression in planarians." Proceedings of the National Academy of Sciences 100.suppl...

  13. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ...from later. The Arabidopsis transformation process The process itself is straightforward. When your agrobacterial...growth on antibiotic-containing media) A similar process occurs in nature; Agrobacterium tDNA insertion ...in hand, so it is pretty important to get this process right the first time. I’ll lay out some guidelines...Agrobacterium strain. Additionally, the transformation process can get a bit messy, so you want to make sure you...begin working in the T2 generation. The selection process itself can affect the growth of resistant plants...

  14. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...CcdB.” Biotechniques. 1996 Aug;21(2):320-3. PubMed PMID: 8862819. 2. Bahassi, EM., et al. “F plasmid CcdB... desired insert should not grow.   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials...still express ccdB and therefore would die. This procedure dramatically reduces the number of clones that...recombinant plasmid and therefore makes the cloning process much more efficient, as one does not have to thoroughly...

  15. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12... before beginning your experiments. Figure 2: Strategy for assembling barcoded combinatorial gRNA...PMID: 25114216. PubMed Central PMCID: PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to... massively parallel combinatorial genetics." Proceedings of the National Academy of Sciences 111.34 (2014... CRISPR-Cas9 screening enabled by CombiGEM." Proceedings of the National Academy of Sciences113.9 (2016...

  16. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...incubation at 37 °C. Wash the membrane for 5–10 minutes 2–3x in fresh wash buffer such as PBS or TBST. Stringent...Purpose Tris HCl 62.5 mM Buffer SDS 2% (w/v) Detergent Β-mercaptoethanol (βME) 100...minutes at 50 °C. Wash the membrane for 5–10 minutes 2–6x in fresh wash buffer such as PBS or TBST. Note .... Dry nitrocellulose membranes are brittle, so proceed with caution. Unlike PVDF, they can be returned...remains, you may want to repeat your stripping procedure, perhaps using longer incubation times, or try...

  17. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...indicated they wanted to see more negative data (Figure 2.) At the same time, when scientists who support open...is not only housed, but encouraged.    Fig. 2: Results from a survey conducted by Addgene in 2021...detailed methods and image descriptions, a minimum n = 2, and well labeled high- and low-magnification images...gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org/10.1089/hgtb.2015.040 Initiative...expression can take months (or longer), yet the process is often not detailed in the primary literature...

  18. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    .... The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose...overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce...Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life...–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours Good separation, quality, and stability ...separation of proteins of different sizes.  Table 2: Recommended gel percentages for various protein sizes...are consistent and stable at room temperature for 1–2 years and are available in formulas specific to your...response over a broad dynamic range that typically spans 2–5 orders of magnitude, which includes a wide dynamic...

  19. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...NGG)  Editing window ~2 nt  85175  pBK-EE-BE3  SpCas9 (NGG)  Editing window ~2 nt  85176  pBK-YE2-...quantum yield of 0.54) and mScarlet-H with M164H (2-fold improvement in photostability compared to mScarlet...SpCas9 base editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated...pBK-YE2-BE3  SpCas9 (NGG)  Editing window ~2 nt  85177  pBK-YEE-BE3  SpCas9 (NGG)  Lower on-target activity...activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity...Ladders Are Awesome Tools: They are comprised of 2 plasmids, pPSU1 and pPSU2, that can be amplified affordably...libraries The Gross Lab at UCSF recently deposited 2 Bacillus subtilis libraries with Addgene – the B. ...

  20. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    Published
    Nov. 1, 2016, 2:30 p.m.
    ...vector:insert ratio is used in the DpnI digestion and 2 µL of digest product are used in the transformation...Biology 38 (2016): 145-154. PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified...in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and vector are first...oldest of these techniques and is a multi-step process consisting of digestion, purification, ligation...techniques with higher efficiency and simpler procedures have been developed. Some of these include, TA...specific sequences. Read on to learn how easy this process is and to get tips for applying it in your own ...Ligation + - Transformation + + The FastCloning process in brief In FastCloning, you PCR both the vector...
Showing: 101 - 120 of 367 results