...: Expand! (Typically by just adding water.)  Step 5: Image   Figure 1: The ExM process allows...applications and there’s just not much to be done about that.  Second, expansion introduces distortions....

...;597(7878):720-725. doi: 10.1038/s41586-021-03886-5. Epub 2021 Sep 6. PMID: 34489594. Eleonora I. Ioannidi.... Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and experts in CRISPR evolution...

...but both rely on the same general strategy. Figure 5: Immunoprecipitation using bead-based antibody capture...are often used in clinical applications. Diabodies - Diabodies contain two Fab fragments, each recognizing...cells isolated from animals immunized as described above are used to create hybridomas, which produce large...different epitope, bound by short peptide linkers. Diabodies can be used for assembling protein nanostructures... Includes: Flow cytometry Read on to learn more about the applications common to each category, including...controls, and any special considerations to think about. For step-by-step instructions of many of these ...protein of interest, as described in the IP section above. Reverse the crosslink, isolate DNA, and perform...

...years and a talk about your research or work will write itself when the time comes. 5. Choose interesting...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...

... the experiment. The second way, it will take you 5 days to complete the experiment, but you’ll only spend...post useful. After a bit of research and thinking about my own processes, I’ve broken down time management...task, you’re actively prioritizing. In my anecdote above, I prioritized splitting cells over meeting up with...was too late. I didn’t spend enough time thinking about the possibilities and how to prioritize them. When...month, or even the year, you should be thinking about two main things: How useful the task is (its utility...plasmids to send to Addgene). Following my advice from above, you should probably take the second option. Even... recommend the second option for the experiment above, but it’s also important not to load yourself up...

...Resources on the Addgene Blog Learn about more tools for reproducibility 5 great apps for lab life Developing...allow you to collaborate within the platform and avoid countless emails. Important collaborative functionalities...of the software. We identified that when talking about ELN costs and limited budgets, “a large percentage...Besides that, academic institutions tend to think about the long term, so they need a solution for which...institution. In the study, respondents were also asked about how important ease of use is for them (if they are...or partners to access your data? If you are collaborating with other labs or clients, you might need share...comments, added files, notes etc. Detailed records about every action performed in the lab (user, date, time...

...naturally occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes...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...

...faster than jGCaMP7 (Figure 5B).   Figure 5: A) GCaMP6, jGCaMP7, and jGCaMP8 viral preps distributed...all, a delivery method. I would be remiss to talk about them without looking at what they’re delivering .... But in the years since, we’ve learned so much about what it takes to make these viral vectors not only...., 2016), video, educational resources, and collaborations, and will continue to do so, in the hopes that...leave a comment; or take our AAV survey. Learn more about GCaMPs and other tools developed through the GENIE...

...the life sciences end up in tenure track position 5 years after receiving their PhDs, still strive to ...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...

...The pCAG-EGxxFP target plasmid contains overlapping 5′ and 3′ EGFP fragments under the ubiquitous CAG promoter...the pX330 plasmids used in the validation assay (above) and it is just as effective (Mashiko et al., 2014...Engineering with CRISPR Validate Your Genome Edit Learn about Non Homologous End Joining and Homology Directed...

... Biology 946 (2013). 1-13. Pubmed PMID: 23179822. 5. Segura, M.M., Kamen, A., Garnier, A. “Downstream ...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...

...templated-repair pathways have these central steps: The 5’ DNA end of the break is resected by nucleases to ...Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips ...

...between 26–45 kb in length. Human adenovirus type 5 (hAdV-C5) can be considered the prototype, although...this introduction gives a sense of what AdVs are about – and we hope that it inspires further use of existing...

...191–197. https://doi.org/10.1016/0378-1119(88)90117-5  Lale, R., Brautaset, T., & Valla, S. (2011). Broad-host-range...plasmids may have high copy numbers and reduced metabolic load (Meyer 2009). Figure 1: Idealized ...

...Molecular Therapy - Methods & Clinical Development 5:106–115 . https://doi.org/10.1016/j.omtm.2017.04.006...therapy. In 2014, RSRT brought together a strong collaboration between two gene therapy labs (Brian Kaspar ...

...Tropism of Lentiviral Vectors through Pseudotyping. CGT 5:387–398 . https://doi.org/10.2174/1566523054546224... also toxic at high concentrations as described above and is inactivated by the complement system in serum...

...working solution by diluting 5 µL of the 300 µM DAPI stock solution into 5 mL PBS. Protect from light. ...coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI) Bovine ... µM DAPI stock solution by diluting 2.1 µL of the 5 mg/mL DAPI solution to 100 µL PBS. Protect from light...well of a 24-well cell culture treated plate. Seed 5 x 10 3 HeLa cells per well. Allow the HeLa cells to... the appropriate container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. Permeabilize cells ...it in an appropriate waste container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. Section 3...it in an appropriate waste container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. Dilute the...

...concentration of 20 μM, then mix: 5 μL Forward oligo 5 μL Reverse oligo 5 μL 10x NEB buffer 2 35 μL ddH 2...cells pUC ori pUC bacterial origin of replication. 5’LTR 5’ long terminal repeat. RRE Rev response element...Forward oligo: 5’ CCGG—21bp sense—CTCGAG—21bp antisense—TTTTTG 3’ Reverse oligo: 5’ AATTCAAAAA—... and a 5kb fragment. 5. Sequence positive clones with pLKO.1 sequencing primer (5’ CAA GGC TGT TAG AGA...-293T cells in 5 mL of media in a 6 cm tissue culture plate. Incubate cells at 37°C, 5% CO 2 overnight...°C. m. Add 5 mL of fresh media containing antibiotics to the cells and incubate at 37°C, 5% CO 2 for 24... Day 5: n. Harvest media from cells and pool with media from Day 4. Spin media at 1,250 rpm for 5 minutes...

...better. The Thomas Kilmann Conflict Model describes 5 basic approaches to conflict resolution based on various...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...

...Biomedicine to 78 undergraduate/graduate students from 5 independent universities in Spain through 22 international...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      ...