We narrowed to 618 results for: abo.2

...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...

... 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...

... 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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...There are approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells...bind the RNA motif will be biotinylated. Read more about RapID in this blog! Find plasmids for RapID EXCELL..., S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki, K., & Roux, K. J. (2016). An improved smaller...

...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...

...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...

...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...

...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...

...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...

...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      ...

...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...

...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...

... 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...