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We narrowed to 675 results for: abo.1

Showing: 181 - 200 of 675 results

  1. Choosing a Good Mentor for Scientists

    Type
    Blog Post
    ...setting out of the lab (coffee, lunch or beer) and in 1:1 conversations. This creates a level of familiarity...Science Mentors series: Part 1: "What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring ... will also serve as a good mentor. To read more about what makes a good mentor, see the previous post ... and personnel management?  If you are thinking about a non-academic path, will the PI support this path...Medical School researcher Connie Cepko to learn about her mentoring style. How can I find out if an advisor...time with the potential advisor and don’t be shy about asking hard questions. Talk to as many lab members... past lab members. Most scientists love to talk about their past lab experiences and will be willing to...

  2. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...acceptor fluorophore. The typical steps in FRET are: 1. Donor fluorophore excitation by absorption of a photon... overlap, the fluorophores must be located within 1-10nm of each other and be oriented appropriately for... from various papers to make the decision easier. 1. Müller, Sara M., et al. "Quantification of Förster...diseases and even assist in drug discovery. References 1. Baird, Geoffrey S., et al. "Circular permutation ...can be done using FRET! Read on to find out more about this amazing imaging technique and find further ...our curated list of biosensors too, as another laboratory may have already created the sensor that you ...ability of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments ...

  3. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ... basic parts of the Cas9 enzyme.   Figure 1: A cartoon depiction of Cas9’s two major lobes, REC...the CRISPR system (Nishimasu et al., 2014). Table 1. Summary of Cas9 domains, the lobe each domain belongs...state (PDB ID 4CMP) with domains colored as in Figure 1. HNH, red; RuvC, orange; CTD, yellow; REC lobe, light... on itself at the bottom of the image. Stem loops 1 and 2 extend horizontally across both lobes. The guide...outside of the protein, spanning both lobes. Stem loop 1, nestled between the REC and CTD domains, is required...Within an interior cavity, Cas9 grips nucleotides 1-10 of the guide RNA, prearranged for base pairing ...343–347. https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales...

  4. Molecular Biology Reference

    Type
    Guide
    ...supE44 thi-1 gyrA96 relA1 tonA JM109 Addgene; Promega e14-(McrA-) recA1 endA1 gyrA96 thi-1 hsdR17(rK- ...galK2 lacY1 proA2 rpsL20 (StrR ) xyl-5 λ– leu mtl-1 Top10 Invitrogen F- mcrA Delta(mrr-hsdRMS-mcrBC) Phi80lacZM15...use, dilute your antibiotic into your LB medium at 1:1,000. For example, to make 100 mL of LB/ampicillin...human genome with restriction enzymes would yield about two million DNA fragments, which is far too many...the molecular biologist’s toolbox. To learn more about different types of cloning methods check out our...restriction enzyme digestion. For more information about restriction enzymes check out NEB's website . Insert...endogenous expression levels). For more information about promoters, both bacterial and eukaryotic, as well...

  5. Expanded Access to Rabies Plasmids

    Type
    Blog Post
    ... export license applications for VSV and SARS-COV-1 plasmids. Export control license process    An export...charged.  Timelines  We have offered VSV and SARS-COV-1 plasmids under this application process since May ...expedite the application process.  More information about export control licenses is available through the...Industry and Security. If you have any questions about this or other Addgene shipping and ordering processes...customer support team at [email protected].  Curious about how rabies plasmids are used in the lab? Check out...

  6. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ... the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the targeted gene have...knock out exon 2 of a given gene.   Figure 1: A knockout targeting vector designed to insert a ... the Neomycin resistance (NeoR) gene between exon 1 and 3 of the targeted gene. Upon recombination, the...inserted GFP is expressed (knocked in). As seen in the above example, you can remove a floxed resistance gene...Get the Basics of Homology Directed Repair Read about Generating Knockouts with CRISPR through Non-Homologous...Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR  ...

  7. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...versatility of the basic antibody structure (Figure 1). Antibodies can be broken up into smaller parts, ...antibodies for a specific purpose. Figure 1: Overview of antibody engineering methods. (A) Basic...,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82, 169, 1)"},"button_font":{"size":19,"styles":{"font-family...:"rgb(255, 255, 255)","rgba":"rgba(255, 255, 255, 1)"}}{% endraw %}{% end_module_attribute %}{% module_attribute...Molecular Biology, 29–62. https://doi.org/10.1007/978-1-0716-2285-8_2 McMahon, C., Baier, A. S., Pascolutti...by PCR. Current Protocols in Molecular Biology, 51(1). https://doi.org/10.1002/0471142727.mb0803s51 Additional...structure-guided directed evolution. The nice thing about these methods is that they all follow the same general...

  8. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... to other technologies like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas... can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath and 42°C ...nearly single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification... Not mentioned in the paper, likely approximately 1-2 hours Equipment needed: Mini PCR machines by DIY-Bio...Sensitivity: – 110 femtomolar  Time: Not mentioned, likely ~1-2 hours Equipment needed: PCR machines/Heat blocks... the Addgene blog Read more about SHERLOCK and DETECTR Learn more about plasmids available for COVID-...method based on CRISPR-Cas12. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.02.29.971127 Rauch...

  9. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...the Brainbow mouse neural labeling system. Brainbow-1 and -2 have been further refined to create Brainbow...subsequent recombinations cannot occur. While the Brainbow-1 system employs Cre-mediated deletion, the design of...antibodies specific to each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained a “default” state; for...dendritic processes not previously visible with Brainbow-1 and -2. The general structure of Brainbow-1.0 is retained...Rheenen J, Simons BD, Clevers H. Cell. 2010 Oct 1;143(1):134-44. PubMed. Drosophila Brainbow: a recombinase-based...integration of multiple copies of the construct (about 8 in Brainbow-1.0 mice.) The combinatorial effect...with the goal of overcoming the limitations listed above. First, they screened a variety of fluorescent proteins...

  10. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ... hybridization experiments. A recent publication [1] from the Sánchez lab describes the plasmids and protocols...deposit) are described in the Sánchez lab publication [1] so that the students performing these experiments...can easily investigate in planarians. References 1. Alice Accorsi, Monique M. Williams, Eric J. Ross,...Proceedings of the National Academy of Sciences 100.suppl 1 (2003): 11861-11865. PubMed PMID: 12917490. PubMed...experiments in the beginning. In most classrooms, the laboratory experience is focused on classical embryology... Biology with BioBuilder 3D Models for learning about CRISPR Resources at Addgene.org Find plasmids ...

  11. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...well, so let’s go over the details step-by-step. 1. Digestion Set up restriction digests for your insert...recipient plasmid : insert ratio” of approximately 1:3. Since the number of base pairs for each varies,...cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent cells...at minimum, two transformations after a ligation: 1. Control Transformation containing the ligation mixture...result in few colonies on both plates (unsuccessful 1), in a vector alone plate with many more colonies ... (post or pre run in the table above, respectively). Some of the above stains require you to visualize...equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid alone...

  12. An Introduction to Adenovirus

    Type
    Blog Post
    ...that the culprit for these unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing...an AdV transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions...certain contexts (Bauzon et al., 2003).        Figure 1: The adenovirus life cycle. Image reused from Kremer... review of the evidence. Emerg Microbes Infect, 8(1), 1679–1687. https://doi.org/10.1080/22221751.2019.1690953...574–580. https://doi.org/10.1038/s41586-023-05949-1. PMID: 36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval...this introduction gives a sense of what AdVs are about – and we hope that it inspires further use of existing...

  13. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Table 1 has a detailed list of the bacterial species these plasmids are compatible with.  Table 1: Non-...high copy numbers and reduced metabolic load (Meyer 2009). Figure 1: Idealized examples of a narrow...for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197. https://doi.org/10.1016/0378-1119(88)90117... N.J.), 765, 327–343. https://doi.org/10.1007/978-1-61779-197-0_19  Meyer, R. (2009). Replication and ...

  14. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...under a wide range of inducer concentrations (0.1-1% L-arabinose) without signs of toxicity or burden ...bacteria have colonized (Fig. 1c).     Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram...ultrasound contrast. In all the cell lines we tested, 1 µg/mL doxycycline was sufficient to saturate expression...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the ... tumors and tumor-homing bacteria. Nat Biotechnol 1–13 (2023) doi:10.1038/s41587-022-01581-y  ...you enjoy playing with our genes. Have fun!    About the authors: Marjorie Buss is a PhD student studying...

  15. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...blog Find our CRISPR blog posts Read blog posts about using Cre-lox Read our genome engineering blog posts... posts Resources on Addgene.org Find more about Cre-lox Read our Genome Engineering Guide Learn more...

  16. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...2021).   Table 1: Properties of modern NIR FPs     NIR FP Ex, nm Em, nm ECa, M-1cm-1 QYb, %   Molecular...brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events via...expression level. State-of-the-art FPs (listed in Table 1) were optimized for efficient binding of endogenous...previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of a probe leads...in eukaryotic cells as an intermediate of heme metabolism (Shemetov et al, 2017). Modern state-of-the-art...NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, performance, and applications...

  17. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ... emailed submission) in the Deposit menu: Figure 1: Begin the deposit process by clicking "Submit Online...the plasmids, or (for deposits submitted after July 1, 2025) after three years on hold, Addgene staff may...cases Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if your deposit falls into...Addgene?" or contact us at [email protected] or +1 (617)-ADDGENE (233-4363) with additional questions...strain. Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if you’d like to distribute...page so that any data you enter is not lost.   Step 1: Sequences, Maps, and Files Please provide any full...a 1.5 mL microfuge tube at a concentration of 0.1–1 µg/µL (or bacterial streaks, if you prefer). If you...

  18. RUBY-Red Siliques

    Type
    Blog Post
    ...in all other plants, anthocyanins.   Figure 1: An adult plant expressing RUBY. Photos by Andrew ... be collected from heterozygous individuals. Only 1/4 of the descendants of the heterozygous plants, therefore... (2020). https://doi.org/10.1038/s41438-020-00390-1 Additional resources on the Addgene blog  CRISPR 101... is difficult for new members of the lab or collaborators unfamiliar with these mutant plants. Using RUBY...resistant disease. (RUBY-red: You can’t beet it!) About the Author  Andrew Willoughby is a graduate student...

  19. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ... vectors are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential...systems have been developed over the years based on HIV-1; however, the 1st-generation vectors are never used...which has been shown to repress expression of HIV-1 LTRs (more information is available here). 2nd vs...Plasmids 101 Learn about genome-wide screening with CRISPR and Lentivirus Read about the ClonTracr Library... the proper cell line is used; several common laboratory cell lines including 293 were immortalized with...

  20. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...require large amounts of high quality DNA. Figure 1 illustrates the three grades of DNA you may need based...their relative quality and quantity.   Figure 1: Plasmid preps fall into different categories based...specific volume ranges of culture as their input (Table 1). When you feel like you need just a bit higher DNA...kit for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher volume can ...beads and a suitable magnet as you scale up. Table 1: Standard Prep Kit Capacities* Prep size Amount...Amount of culture Approximate DNA yield** Miniprep 1–5 mL 5–50 µg Midiprep 25–100 mL 20–200 µg Maxiprep...columns we’ve been talking about!) are a significant source of waste in laboratories. To help combat this, ...
Showing: 181 - 200 of 675 results