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We narrowed to 604 results for: abo.2

Showing: 521 - 560 of 604 results

  1. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...of these issues are not particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges...have been written about CRISPR alone, and Zimmer himself has written entire books about optogenetics. Defining... been thinking about a long time. For the scientists who like to learn everything about one specific topic...wisely makes no such attempt. He is transparent about his own biases--many of which will be shared by ... a massive, expensive project that required collaboration from scientists around the world. The paper ...As a scientist who enjoys learning a little bit about many different things (part of the reason Addgene...

  2. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...PMID: 24901218. PubMed Central PMCID: PMC4047077. 2. Pagliara, Patrizia, et al. "Ostreopsis cf. ovata ...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

  3. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...Reports 9: https://doi.org/10.1038/s41598-019-41088-2 Magnus C, Lee P, Bonaventura J, Zemla R, Gomez J, ...posts Read blog posts about neuroscience Resources on addgene.org Learn more about chemogenetics in our...as a DREADD agonist due to its ability to be metabolized into clozapine (CLZ). While some scientists argue...

  4. Of Myc and Men

    Type
    Blog Post
    ...promising monoclonal antibody with high specificity. (2) Narrow down the specific epitope that your antibody...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take...times of the myc tag.  Origins of protein tags For about as long as the disciplines have existed, cell and...examples of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable ...

  5. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ... replication competent virus [usually HIV-1]; and 2) the potential for oncogenesis through insertional...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...

  6. Rabies and Neuronal Tracing

    Type
    Blog Post
    ... 2008 Dec; 18(6): 617–623. Pubmed PMID 19349161.  2. Deshpande A, Bergami M, Ghanem A, Conzelmann KK, ...connected to the neck. Shortly after, we learned about organs and the jobs they perform. This became foundational...part of the answer to many biological questions about the brain. To address this, molecular tools to map...

  7. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...integrates into the trnS-leu2 locus.   Figure 2: Introducing Mobile-CRISPRi into bacteria. Top: A ...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko...

  8. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...PMID: 25538079. PubMed Central PMCID: PMC4353587. 2. Stephen, Andrew G., et al. "Dragging ras back in ... cancer are yielding promising results in the laboratory, and Addgene is accelerating this research by...within that pathway that are of interest to their laboratory. A single mouse click on a gene of interest will...

  9. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...that view (Callahan et al., 2022).     Fig. 2: Students in an MDH CURE replaced select residues ...in a year-long biochemistry laboratory : Part II – research-based laboratory – a semester-long research...are often associated with rote memorization of metabolic pathways (precisely the type of exercise Einstein...had the opportunity to talk with Provost recently about his development of what is now commonly termed a...into one of my own courses, I was excited to hear about how he developed his program. Initially, Provost...confidence in their research capabilities and laboratory and critical thinking skills (Knutson et al.,...importance of acknowledging student apprehensions about “failure” (protocols not working, hypotheses being...

  10. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...that numbers are not switched around or misread.  2. Track the container - It is just as important to ...is the Laboratory Operations Manager at Addgene.  In addition to helping with daily laboratory operations...through barcoded tubes that are scanned into our Laboratory Information Management System (LIMS), a database...available for distribution!    Check Out How We Think about Developing Lab Management Software    The plasmid's...the correct sample. Tracking samples - it's all about location, location, location There are many different...of these samples have the 2D barcodes mentioned above. These barcodes identify what type of sample is ...every barcode on every tube contains information about that tube’s appropriate location. The barcode also...

  11. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...You’ve probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering... to devise an experiment that can tell you more about its function.  Resources and references Resources...

  12. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ...would help them grasp some of the key concepts (Fig. 2). One thing I will likely change for next time is ...wonderful tangents. Some groups asked about vaccines, others were curious about the agricultural repercussions...roles and responsibilities, I have to think a lot about how to balance those roles. I’m a father of three...community. This past year, I spent some time thinking about how I could combine all of these roles by reaching...guest presentations. If you’re a scientist thinking about reaching out to your community’s schools, here are...accountants running our math tables!  Be realistic about what you can teach  5-10 year olds in a single evening...DNA, protein, plasmid, gene, etc. Though I talked about CRISPR’s molecular mechanism, I was careful to keep...

  13. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...externally fertilized, transparent progeny (Figure 2). These features give it a leg up (or a fin up?) over...for gene manipulation together with information about the genome are powerful resources for investigating...zebrafish genome was published in 2013, revealing that about 70% of human genes have a counterpart in zebrafish...networks that regulate processes from development to metabolism to fertility. Several zebrafish-specific gene...gene expression databases that provide information about when and where genes are active in developing embryos...genome sequences become available, new hypotheses about specific genetic interactions arise and tools to...genome, their long three-month generation time and labor-intensive maintenance compared to invertebrate model...

  14. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ... the Cas machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA...dependencies associated with each ancestry group. Since about 90% of the cell lines used in DepMap CRISPR screens...

  15. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ...interesting things to do, feel free to crowdsource! 2. Free food generally gets people to show up If you...early employees spent this remaining time by elaborately pranking each other, dressing in costume, and...frequently ask the question, “What’s your favorite part about working at Addgene?” Most Addgenies reply with an... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...rose-colored-glasses-esque picture of how everyone is kind and collaborative and all we do is have fun all day long, whistling...megaphone, or your company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring...company.        Additional Resources Read other posts about life at Addgene!Read more of our careers posts! ...

  16. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...experience and to build upon your existing skills.  2. Find your niche Identify and develop an area of expertise...on becoming grant writers. When we asked others about how they became grant professionals, mid- to late-career...late-career grant professionals have similar stories about a time when their passion for an organization or... logic models, established partnerships and collaborations, and commitments to and proof of compliance...grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers... skills & passion  Skills such as research, collaboration across multiple disciplines, writing, project...development strategy builds on the steps listed above and formalizes your development. You may consider...

  17. The Challenges of Cell Culture

    Type
    Blog Post
    ...Henrietta Lacks. New York: Crown Publishers, 2010. 2. Gold, Michael. "A conspiracy of cells." State University...Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about the Dangers...Misidentified or cross-contaminated cells have been known about since 1966 when Stanley Gartler reported that most...there is a loss of reputation that can impede collaborations, especially with industry. Today we also have...The LGC Group also provides standards and has laboratories to carry our independent quality control for...culture contamination." Archives of pathology & laboratory medicine 133.9 (2009): 1463-1467. PubMed PMID...

  18. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...most important” tasks my schedule. This was bad for 2 reasons. First, it soon became obvious that I didn... think much about management skills during your scientific training. I never thought about management ...open mind. First let’s clear up a misconception about being a manager: yes, being a manager means being...you should be doing : writing a grant, thinking about the next project, reaching out to other teams, expanding...buffer them from unnecessary pressure “coming from above”, which is when other skills such as negotiation...

  19. AAV ddPCR Titration

    Type
    Protocol
    ... 95 10 2 1 Denaturation 95 0.5 2 50 Annealing/Extension 60 1 2 50 Signal Stabilization 98 10 2 1 Hold ...should decrease by a factor of 2 across the dilutions. In the example below, 2-fold serial dilutions of a ... considered biosafety level 1 but may require BSL-2 handling depending on the insert. Please ensure that...single channel pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...20X): 5 µL in 95 µL 1X PCR buffer (1:20) Dilution 2 (20X): 5 µL in 95 µL 1X PCR buffer (1:400) Dilution... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction...Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft software...

  20. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 Watch the Video! Water...BSL-1 and BSL-2 Labs Learn how to best protect yourself when working in BSL-1 and BSL-2 labs Watch the...maintenance and use Watch the Video! Pipetting Learn about selecting the correct pipettor and pipette tip, ...the pipette Watch the Video! Centrifugation Learn about selecting and using a centrifuge to separate different... a liquid sample Using a Light Microscope Learn about the parts of a light microscope and its use Weighing...Weighing Reagents Learn how to weigh laboratory materials on a balance Basic Molecular Biology Name Description...

  21. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis...genes above with your gene or genes of interest. Some of the plasmids listed in the table above (and their... an internal region of the mRNA. In the figure above, you can see that the IRES element acts as another... first discovered in picornaviruses, are short (about 20 amino acids) and produce equimolar levels of ...

  22. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ... PMID: 25184501. PubMed Central PMCID: MC4262738. 2. Doench, John G., et al. "Optimized sgRNA design to...Read this blog post about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources on...libraries for the mouse genome! Other Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...

  23. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work...worry less about how they will handle an emergency and this will make you much more confident about their ...

  24. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms have...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...Effects on transcription (remember, it’s not all about translation - e.g. codon choice may interrupt transcription...

  25. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...agarose concentrations (e.g., 2 g of agarose in 100 mL of TAE will make a 2% gel). Mix agarose powder with...concentration of approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr...gels are commonly used in concentrations of 0.7% to 2% depending on the size of bands needed to be separated...allows you to gauge how far the DNA has migrated; 2) it contains a high percentage of glycerol that increases...cool down to about 50 °C (about when you can comfortably keep your hand on the flask), about 5 mins. Optional... of the tip of the pipette into the buffer just above the well. Very slowly and steadily, push the sample... of the way down the gel. A typical run time is about 1-1.5 hours, depending on the gel concentration ...

  26. Design Tips for Prime Editing

    Type
    Blog Post
    ...inhibit editing (Anzalone et al., 2019). Figure 2: Starting lengths for optimizing your pegRNA primer... primer binding site, starting with a length of about 13 nt (Anzalone et al., 2019). Primer binding sites...of reverse transcriptase template, starting with about 10–16 nucleotides. For longer templates, it is even.... Test multiple nick sites, starting with sites about 50 bp upstream and downstream from the prime editing...

  27. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...Hoogsten base pairs and wobble pairs.  Figure 2: (A) The codon wheel depicts the 64 codons that encodes...usage bias Read more about plasmids in our Plasmids 101 blog series Learn more about trends in synthetic... out Addgene's Molecular Biology Reference Read about genetic code expansion Browse our plasmid collection...

  28. Hot Plasmids - August 2020

    Type
    Blog Post
    ...based antiviral method to degrade RNA from SARS-CoV-2 sequences and live influenza A virus. It was demonstrated... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's... a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing effort to improve genetically..., that are optimized for excitation wavelengths above 1,000 nm and enable two-color imaging with red fluorescence...

  29. Bioinformatics at Addgene

    Type
    Blog Post
    ...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...much information as possible from depositing laboratories and associated publications, but our quality...the process. Another design choice we appreciated about Nextflow was how effortless it was for us to switch...interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our pipeline...

  30. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...Generating a transgenic plant takes even longer, is more labor intensive, inefficient, and often not reproducible... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo system...

  31. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ...) or the American Chemical Society (ACS) are just 2 examples of organizations that hold large, annual ...or postdoc forever, and you need to be thinking about what comes next. To that end, one of the most important...personal connection; the importance of which will be elaborated on later. As for what job fairs to consider, ...the specific position he or she originally called about, you can keep in touch and reach back out when you...

  32. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct... more about CcdB, the toxic key to efficient cloning Read all Plasmids 101 blog posts Read about knockout...with the other toxin-antitoxin systems described above, one part of the pair is more stable than the other...

  33. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...rules are: (1) don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same...cycling. The Pros and Cons Researchers used the tools above to complete a 52-piece assembly of a 40 kb T7 phage... help from DAD. The 35-piece assembly mentioned above had a remarkably high 71% fidelity, while the 52...

  34. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...section 1, step 2 is a suggested starting point. If your unknown sample’s absorbance falls above the range ...vapors. Workflow Timeline Day 1: Antigen Coating Day 2: Blocking Day 3: Primary antibody incubation Day 4...with 96-well plates 1–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...serial dilutions of the purified antigen as follows: 2 ng/µL : Add 100 µL of 20 ng/µL stock into 900 µL PBS...microfuge tube and vortex. 1 ng/µL : Add 450 µL of 2 ng/µL stock into 450 uL PBS in a microfuge tube and...37 °C for 30 min , or overnight at 4 °C . Section 2: Block the plate Prepare the wash buffer (0.05% Tween...incubate on a microplate shaker set to 400 rpm for 2 h at room temperature or overnight at 4 °C . Section...

  35. Kit Free RNA Extraction

    Type
    Protocol
    ...RNAzol®, QIAzol® (for Protocol Option #2) Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl... reagent section above for the recipe). If using TRIzol®, jump down to the Option #2 - TRIzol® Protocol....5% (wt/vol) N-laurosylsarcosine (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as...following sequentially to 1 mL of lysate: Add 0.1 mL of 2 M sodium acetate (pH 4.0), mix thoroughly by inversion...aliquots of it and storing those in -80°C. Option #2 - TRIzol® Protocol Homogenize or lyse tissues or cells...by hand for 10 seconds. Incubate the sample(s) for 2-3 minutes on ice and centrifuge for 15 minutes at ... working with the volatile reagents in the list above. Procedure Option #1 - Solution D Protocol Before...

  36. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...rate of 10-55% - the PASTE system.     Fig. 2: Schematic of programmable gene insertion with PASTE.... Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and experts in CRISPR evolution...

  37. Hot Plasmids - November 2023

    Type
    Blog Post
    ... vivo and in vitro applications.   Figure 2: Modular features of DIRECTED particles where the ...section of the Addgene antibody page, and learn more about the whole-brain reference atlas efforts, including...

  38. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...replicate, then purify the DNA.     Figure 2: A pooled library page with protocols boxed in red...ordering a pre-made library, you don’t need to worry about amplifying and producing lentivirus. But pre-made...

  39. Immunocytochemistry

    Type
    Protocol
    ...Fluorescent microscope 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI) Bovine serum albumin (BSA) Triton...grow to the desired density before labeling. Section 2: Fixing and permeabilizing cells Gently aspirate the...of the diluted antibody to the wells and incubate 2 h at RT . Remove the primary antibody and dispose ...water Microscope slide Anti-fade mounting medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes...paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of waste in the...remove the coverslip. Blot the coverslip with a laboratory wipe to remove excess liquid. Add 1 drop of anti-fade...

  40. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... glacial acetic acid 57 mL of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures from single...Resuspension buffer Denaturing solution Renaturing solution 2 mg/mL RNase A TE or water-saturated phenol-chloroform...100 μL of cold Solution I. Vortex the solution for 2 min or until all bacteria are fully resuspended. Add...pipetting or carefully pouring. Optional: Add 5 μL of 2 mg/mL RNase A to the supernatant in the new tube and... to the recovered aqueous DNA layer. Repeat steps 2-4. Note: Phenol-chloroform is a hazardous waste - ...: Ethanol Precipitation To your DNA solution, add 2-2.5 volumes 95% or 100% ethanol and 1/10 volume of... supernatant out of the tube if you are worried about losing the pellet. Dry with vacuum or by inverting...
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