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We narrowed to 604 results for: abo.2

Showing: 561 - 600 of 604 results

  1. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ... Watch Joanne give her "Not" Networking 101 Talk 2. Tailor every talk to the occasion Don’t just plan...interesting content Talk about something you know a lot about and that you care about. Don’t try to present...engaging and memorable. If you read my story above about the presentation faux pas, you are more likely...the room, so it is a good idea to start thinking about public speaking early and often. Two of my jobs ...dynamite presentation?  To that end, let’s chat about giving science talks and how to make them serve ...other people give great talks We are not talking about lab meeting here. Lifeless presentation of experimental...You want to give the audience something to think about, argue with, or learn from. Some of the best talks...

  2. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...approximately 50–60% confluent. For 293T cells this is about 2 × 10 6 cells in a 10 cm dish. Each 10 cm dish should...Day 1: Seed individual cells in a 96-well plate Day 2–14: Monitor cells for growth and expand cells Day ...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...channel pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller Hazardous waste container...the highest or lowest transgene expression ( Figure 2 ). Sample Data Figure 1: Generation of monoclonal ...inferred by the differences in colony size. Figure 2: Cas9 expression in monoclonal cell lines generated...cell solution into the conditioned medium prepared above to make a new cell solution at a concentration of...

  3. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...questions! 2. The r/Science subreddit is well moderated. One of our original worries about sharing science...not too worried about selling yourself and really do just want to get information about yourself and your...where users can participate in forum discussions about a wide variety of topics. Individual forums are ...where users can ask designated scientists anything about a particular topic. Given Joanne’s expertise in...answering questions. r/Science has pretty strict rules about not being too sales-y in posts which falls in line...any scientists to refer to if they have questions about science careers, women in STEM, work-life negotiation...providing us with new questions and topics to write about in the future. The AMA was also cataloged for permanent...

  4. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control...Spinach-based riboswitches can be used as stable metabolite sensors for imaging in live cells. By placing...placing Spinach downstream of a riboswitch, metabolite binding to the riboswitch instead activates Spinach ...Dimerization of Broccoli increases its fluorescence by about 1.8-fold, and may be a better choice for imaging...modification that reduces nonspecific binding with RNA by about 7x). Mango fluoresces nearly as brightly as Spinach...Additional resources on the Addgene blog Learn more about aptamer fluorophores Investigate other fluorescent...

  5. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...to the break room for a sip of water.  Fig 2: Blugene likes to spark creativity in out on the open...brainstorming sessions.  Allow the group time to think about the problem before you meet. This holds the dual... meetings run smoother when people have thought about the issue ahead of time. Encourage dissent. The ... decisions while encouraging the group to think about alternative options. When a devil’s advocate is ...a few minutes every month dedicated to thinking about the future will improve your ability to innovate...to be able to continue our experiments. Thinking about these plans before you’ve run out of consumables...

  6. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ... plate. Incubate plates at 37 ℃ for 18 hours. Day 2 Observe plates for colony formation. Shown below are...individual colonies and effective selection. 150 µL of 2 mg/mL Carbenicillin plated over-agar Plate shows less...individual colonies with smaller size than the 1 mg/mL and 2 mg/mL plates and effective selection. Selection Curve...coli after Over-Agar Plating of Carbenicillin. The above graph displays the stock concentration of Carbenicillin...concentrations that will work for this assay, and the above result represents a single experiment. For publishable...

  7. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...Postdoctoral Associations around Boston - From Panel Session 2: Defining the Postdoc: An Institutional Perspective...compile more comprehensive demographic information about the postdoc work force, problems start with the ...by Paula Stephan is $40,000) Increase Knowledge about Postdoc and Graduate Student Career Outcomes University...through many iterations read by your PI, your collaborators, others you know in the field, and even people...important questions that still need to be answered about the makeup and welfare of the many researchers out...

  8. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression...How does inducible expression work? As mentioned above, many expression plasmids utilize inducible promoters...see protein overexpression? The strains described above should generate sufficient expression levels for...may help with solubility issues. We want to hear about your favorite protein expression tips and tricks...

  9. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...positive cells (Martin et al., 2019). Figure 2: Editing efficiencies using base editing reporters...for base-editing activity The reporters mentioned above all rely on GFP fluorescence and cell sorting using... Additional resources on the Addgene blog Read about other CRISPR reporters Browse our CRISPR blog posts...

  10. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...Nature 339 (1989). 487–488. PubMed PMID: 2725683. 2. Miller, C.J., Kassem H.S., Pepper S.D., Hey Y., Ward...Lentiviruses are a powerful laboratory tool often employed to establish cell lines that stably express...Get Tips for Titering Your Lentivirus Learn All About Viral Vector Elements Learn How to Do Genome-Wide...

  11. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...mL glass bottle: 4 g NaCl 4 g Tryptone 2 g Yeast Extract and dH 2 O to 400 mL Note: If your lab has pre-mixed... to do a mini-prep you will usually want to start 2 mL in a falcon tube, but for larger preps you might...might want to use as much as a liter of LB in a 2 L Erlenmeyer flask. Using a sterile pipette tip or toothpick...suggested amount, instead of the other dry ingredients above. Media without growth (top) and with growth (bottom...

  13. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...PubMed PMID: 3459152. PubMed Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl... Brush up on Your Cloning Techniques Learn More about Plasmids in Our Plasmids 101 Series Use CRISPR to...

  14. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...start selecting and testing systemic capsids! Figure 2 contains a decision tree for selecting a capsid, while... highlighted below the table.     Figure 2: This decision tree can help guide your choice on ...transfer to the adult brain. Nature Biotechnology, 34(2), 204–209. https://doi.org/10.1038/nbt.3440 Goertsen... for their use. Then, we’ll provide information about commonly used and emerging capsids to help you decide...our Caltech Systemic Capsids page to learn more about that collection. Table 1: Systemic capsid plasmids...slices Several of the systemic capsids mentioned above have also been tested on ex vivo brain slices obtained...experiments and Addgene’s AAV Data Hub to learn more about the capsid you’re interested in. Then request your...

  15. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...Workflow Timeline Day 0: Seed and transduce Cells Day 2–3 (am): Remove media, replace with fresh media containing...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... 200–1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container...cell death, the cell media should be changed every 2–3 days to maintain the dose of antibiotic, which may...confluent 10 cm dish can be expanded into two 75 cm 2 flasks, etc. Pro-Tip This selection method results...cells in the untransduced well (0 µL lentivirus, above) are dying. Perform regular media changes and monitor...

  16. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...rely on what I know about those particular teams at that given time. I think about how previous iterations...disagreements. In fact, conflict resolution isn’t just about resolving conflicts between other people, but resolving...normally get along well with each other disagree about a specific decision, direction, or goal. Resolving...of lapsed communication. Making some assumptions about how something should be communicated is necessary...confusion around... Ownership: Misunderstandings about who is responsible for what tasks or projects is...conflicts.  Transitioning a conflict into a collaboration may not be the easiest resolution to accomplish...

  17. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ...cooling to room temperature (~45 minutes). Method #2 Place mixed oligos in a PCR tube. Place tube in a ...a thermocycler programmed to start at 95°C for 2 minutes. Then, gradually cool to 25°C over 45 minutes...vector with 0.75-6 ng of annealed oligos). Transform 2-3μL into your favorite competent bacteria and plate...water and quantify the concentration (should be about 8ng/μl). Mix the annealed oligos with cut vector...

  18. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    ...15.4%) 46 (59%) of these students were in the last 2 years of their degree. Students filled in a pre-program...USA-Europe professional collaborations, setting up bases and contacts with Spanish laboratories to start working...where individuals can exchange ideas, network, collaborate, and innovate internationally. Why an International...needs, and came from the universities mentioned above. The initial IMP cohort of 78 mentees included: ...    Other Resources Visit the IMP website Learn about the Mentor Circle Program at Brigham and Women's...Networking for Scientists Series Read our Blog Post about Choosing a Mentor      ...

  19. Five Popular Model Organisms

    Type
    Blog Post
    ...variety of environmental conditions, and double every 2 hours. Yeast are also the first eukaryotic genome ...the time between being born and giving birth - of about 10 weeks. Adult mice reproduce as often as every...are great model organism not only for the reasons above but because they actually share many biological ...cells, yeast DNA is packaged into chromosomes and about 23% of yeast genes have a counterpart in humans ... become an important model organism. They share about 70% of their genes with humans and 85% of human ...in the popular model organisms series, and read about new systems being developed for emerging model organisms...

  20. A Control for All Seasons

    Type
    Blog Post
    ...as the protocol remains the same.    Figure 2: Microscopy-based antibody experiments should include...subjected to the experimental condition. In the example above, a negative control could be a third group of mice...in the experimental protocol. In the experiment above, you could repeat the tau western blot several times...Baseline secondary signaling levels In addition to the above-mentioned safeguards, you’ll need to test the secondary...Flow cytometry In addition to the controls listed above, viability and compensation controls are recommended...technique before starting. For more information about antibodies and antibody-based experiments, check...

  21. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...uses of biotinylation in molecular biology! Learn about in vivo biotinylation of bacterial fusion proteins...covalently attached to the active site of key metabolic enzymes including biotin carboxylase and decarboxylases... organic acids to produce important cellular metabolites. Biotin acts as a carboxyl carrier from one compound...such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction. Biotinylation is the...

  22. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... more from the authors in this blog post. Figure 2: iGluSnFR3 displays enhanced brightness, expression...screen (Koveal et al. 2022). Lactate is a key metabolite in glycolysis, so this sensor will be a valuable...valuable tool to study cellular physiology and metabolism under various conditions. LiLac has improved response...going viral! In addition to the viral preps noted above for iGluSnFR3 and JEDI-2P, several biosensors already...innovation next year! Did we miss any? Let us know about your favorite new biosensor plasmids in the comments...

  23. Molecular Biology Reference

    Type
    Guide
    ...indicated, the antibiotic powder can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions...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...can be replicated easily and efficiently in a laboratory setting. Stable - Plasmids are stable long-term...codon. Below you will find helpful resource tables about the genetic code. This table includes the nucleotide...

  24. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...In our last post, we talked about the first base transversion editors: CGBEs, or C → G Base Editors. ...apyrimidinic sites and abbreviated as AP sites. A collaborative team from the David Liu and Dali Li labs reported... CRISPR 101: Cytosine Transversion Editors Read about four base editor reporters Resources on Addgene.org...

  25. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...two visible blue or magenta chromoproteins (Figure 2) (Liljeruhm et al., 2018). This system is a nice confirmation... a sucrase that converts sucrose into a toxic metabolite. Cells that have undergone a double crossover...confirmation of successful single crossover – after about 48 hours, the resulting colonies turn subtly blue...blog posts Find our CRISPR blog posts Learn more about Lambda Red recombineering Resources on Addgene....

  26. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ... iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells vs. iRFP713, % d ...in eukaryotic cells as an intermediate of heme metabolism (Shemetov et al, 2017). Modern state-of-the-art...reporters and biosensors NIR reporters can tell us about cellular events via  fluorescence changes occurring...NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, performance, and applications...

  27. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ... Methods 2009; 6, 343-345. PubMed PMID: 19363495. 2. Wang JW, Wang A, Li K, Wang B, Jin S, Reiser M, Lockey...the annealed and filled-in gaps The great part about this mix of enzymes is that they can all work at...the standard Gibson assembly techniques described above, this resulted in the direct and seamless cloning...Blog Browse All of Our Plasmids 101 Posts Read About Other Cloning Techniques Additional Resources on...

  28. Protocol - How to Streak a Plate

    Type
    Protocol
    ...streak #2. Using a third sterile pipette tip, toothpick, or sterilized loop, drag through streak #2 and spread...resistance and your initials. Labeling within a laboratory setting is important for organization, and it...a section of the plate, as shown in the diagram above, to create streak #1. Pro-Tips Hold your tooth pick...

  29. CRISPR Library Amplification

    Type
    Protocol
    ... recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be performed at the... day to ensure that growth times are limited. Day 2: Harvest cells and purify DNA (Estimated time 3-4 ... Tubes should contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations...has been absorbed by the agar. This usually takes 1-2 minutes. Critical Be careful not to rip or shred the...incubation if needed. Place 100 mL sterile LB at 4 ℃. Day 2 (morning) Before beginning, prechill at least four...pellet. The total weight of each pellet should be ~1-2 g. Pro-Tip Make sure to weigh the empty tube beforehand...require modifications dictated by the originating laboratory for optimal results. If you obtained the pooled...

  30. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...s) Buffer BSA (if recommended by manufacturer) dH 2 O up to total volume Pro-Tips The amount of DNA that... 10x Buffer 3 µL 10x BSA (if recommended) x µL dH 2 O (to bring total volume to 30µL) The amount of restriction...from 45 mins to overnight. For diagnostic digests, 1-2 hours is often sufficient. For digests with >1 µg ...more at (Link opens in a new window) NEB's website about star activity . If you are digesting a large number...

  32. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...excise the HIV-1 genome from every infected cell and 2. prevent reinfection of those cells. Kaminski et al...reservoirs, chronic inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune...

  34. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... from damage (Abe & Lim, 2024).    Figure 2: A) When proteins (blue) interact with the air-water...transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration with the Vallabh/Minikel Lab, the Weissman Lab...

  35. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...signal separately for each target.   Figure 2: Examples of simultaneous multiplex IF labeling of...to label the rat brain sample shown in Figure 2A above, Andrews et al. used the following combination of...

  36. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    ...Bachelor's degree Generally a short-term position (1-2 years) Often a gap-year position taken to transition...dig deep Talk to your professors about their research, and learn about what makes them excited to do what...sciences), people conduct research to answer questions about how the world works. As scientists, we probe beyond...curious person. I like to ask questions and think about the best way to answer those questions. I’ve met...enjoy, is worthwhile and can help us make decisions about what we want to do in the future. How do I get started...sense of what kind of research you would be excited about. I know quite a few friends who opted to join labs... a lot out there worth exploring! Being excited about the science allows you to ask more interesting questions...

  37. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...ask new and interesting questions in your own lab. 2. Seek multiple mentors during your postdoc At the ...Nature and Science, for example). This blog post is about how to get the most out of your postdoctoral training...your new model?  PFs need to be regularly thinking about manuscripts. The other items will follow. If you...this is a strong indicator that this person cares about your science and your development. Taken together...Researchers face a fundamental conflict. Science is about exploration, discovering new information. There ...many PFs are reluctant to give talks or posters about their research. They dread giving a departmental...manuscript reviews, etc. You'll also meet future collaborators and get access to unpublished information and...

  38. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. ...is referred to as an antigen. You can learn more about antibodies in our Antibodies 101 animation, What... expressing this protein. Read on to learn more about antibodies and how to use them in the lab! Parts...lots is a big concern. You can find more details about polyclonal antibodies on our Antibodies 101: Polyclonal...not necessarily what you are getting. Learn more about monoclonal antibodies here. Recombinant antibodies... as well as compare between samples. Learn more about flow cytometry. Cell and tissue labeling Antibodies...Additional resources on the Addgene blog: Read about another reagent sharing platform for antibodies,...

  39. Pipetting Protocol

    Type
    Protocol
    ...right. Pipette Dispense Volume P2 0.2 to 2 µL P10 1 to 10 µL P20 2 to 20 µL P100 10 to 100 µL P200 20 to ...Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and how to handle...liquid (think: 0.1 µL to 1 mL). When working in a laboratory, properly dispensing liquid ensures the accuracy...100 from top to bottom (as shown in the picture above), while 650 µL would read as 065. P200: The first...100 from top to bottom (as shown in the picture above), while 95 µL would read as 095. P20: The first ...10 µL would read as 100 (as shown in the picture above), while 2.2 µL would read as 022. P2: The first ...1 µL would read as 100 (as shown in the picture above), while 0.5 µL would read as 050. Each volume display...

  40. Finding Your Perfect Job After University

    Type
    Blog Post
    ...experience in cancer research After graduating with a 2:1 BSc in Molecular Biology (roughly a B average in...really passionate about during my degree. I loved parasitology. However, when I enquired about roles in this...with an overview of my experience and how I felt about the different positions to enable them to decide...
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