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Showing: 461 - 480 of 639 results
  1. Plasmids 101: NGS Quality Control for Pooled Libraries

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    Blog Post
    ...created for the Brunello library is shown in Figure 2. You can use a small PCR product because the plasmids...next-generation sequencing (NGS). For gRNA libraries, you’ll design primers to create 200-300 bp sequencing products...
  2. Resume Writing for Non-academic Science Careers

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    Blog Post
    ... remember that a resume is typically no more than 2 pages long. Write your resume for the job posting ...University of British Columbia, Student Services - includes videos on general resume writing, as well as additional...
  3. Viral Vectors 101: The AAV Retrograde Serotype

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    Blog Post
    ...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected...trillion connections (synapses.)  In this post we’ll describe the AAV retrograde serotype (AAVrg) and how it... this can cause challenges for the experimental design as these vectors can exhibit spurious expression...efficiency can have a major impact on experimental design and interpretation of results. Take extra care ...References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant permits efficient retrograde access...
  4. Plasmids 101: Methylation and Restriction Enzymes

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    Blog Post
    ...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control... NEB's Resource describing Dam and Dcm Methylases Discovery of DpnI Review describing dam-/dcm- E. coli...mutagenesis. During this process, incorporation of a desired mutation into your plasmid of interest by PCR generates...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which...
  5. Summer SciComm Series: A PhD in Science Communication

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    Blog Post
    ...perceived climate change risks. Nature Climate Change, 2(10), 732-735. doi:10.1038/nclimate1547 Kunda, Z. (...abstracts were driven by the quality of the study design or by their preconceived ideas about the controversial...transmission and how different fabrics and mask designs could work to slow the spread of illness. I later...Series: Masters of SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication...
  6. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...responsible for that differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced...transcription factor library The human genome encodes approximately 1,800 TFs, with >3,500 unique isoforms... and screening mechanisms, although the unique barcodes for each TF help streamline identification. While...a disease in a model system. TF screens can be designed to identify custom TF programs to acutely initiate...
  7. Communicating Your Science With Help From ComSciCon

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    Blog Post
    ...aspects of the work you presented may also be unclear. 2. Make the audience your primary focus When writing... craft sentences for abstracts and presentation slides, learn from audiences as they do science outreach...about whether the same principle applies to other slides in your deck.  If someone asks a question about...of a new science communication initiative that provides yet more scientists with opportunities to practice...
  8. Antibody Validation for Flow Cytometry

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    Blog Post
    ...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach...see: Uhlen et al., 2016). In this post, we will describe the approaches that can be used to determine selectivity...intended application — here, flow cytometry — is the desired target, using one or more of the approaches outlined...interpreting your data therefore necessitates careful design of RNAi sequences, confirmation of RNA knockdown...the target at different levels, it is possible to design experiments where the performance of the antibody...We find that cell tracker dyes are helpful in designing such panels, so that pre-stained cell lines can...
  9. Plasmids 101: Biotinylation

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    Blog Post
    ...Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...26224555. PubMed Central PMCID: PMC4884002. 10. Boer, E. de, Rodriguez, P., Bonte, E., Krijgsveld, J., Katsantoni...Northern blotting Cell surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated...your downstream application. Biotin labelled oligonucleotides can be used in a multitude of applications...commonly used this method to attach proteins or polypeptides to a solid substrate (Holden and Cremer, 2003...
  10. Building Global Connections with the International Mentorship Program USA-EUROPE

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    Blog Post
    ...University of Oviedo, the European University Miguel de Cervantes, the University of Navarra, and the University...15.4%) 46 (59%) of these students were in the last 2 years of their degree. Students filled in a pre-program...together with the universities, a bright future. IMP provides matched one-on-one international mentorship, with...recommended, a mentor with international experience provides his/her mentees with a broader, first-hand vision...
  11. K. phaffii: Rising to the Occasion

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    Blog Post
    ...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...biomass production techniques in the following decades, Phillips made the decision to release the K. phaffii...of K. phaffii lacks certain terminal mannose glycopeptides (which most other yeast possess) that are problematic...
  12. Plasmids 101: Mammalian Vectors

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    Blog Post
    ... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK...mammalian plasmid components, it may be useful to describe the means of introducing genetic material (such...the plasmid in the same way. Selection methods, described below, are typically employed when creating stable...
  13. AAV Q&A with Tim Miles

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    Blog Post
    ...retransduce a model organism such as mice, e.g. first at 2 months, then at 4 months? Or do antibodies become ...1x1012 vg/mouse is more standard) and reflects a desire for very high MOI on top of very high coverage....researchers have used the flexibility of AAV genome design in remarkably creative ways. The continued development...
  14. Hot Plasmids - May 2022

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    Blog Post
    ... those that also infect the liver.    Fig. 2: Summary of the generational evolution of the AAV ... error rate for long sequence knock-ins Visual barcodes for live cell clonal multiplexing Improving ...nanobody-GFP interaction). The modularity of the system provides researchers better flexibility and makes the approach...other experiments. And, while the BLITZ system was designed and optimized with zebrafish in mind, it can easily...https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal multiplexing by: Rachel Leeson...Recently, Ravid Straussman’s lab has developed visual barcodes for clonal multiplexing of cells in live imaging...peptide. By combining three different localization peptides with four different fluorescent proteins, and ...
  15. Hot Plasmids - February 2022

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    Blog Post
    ... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While two... two guides require two arrays for Cas9, two guides can be streamlined into a single array for Cas12a....applications. This set of new anti-FLAG frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag ...target multiple genes within a single cassette provides increased simplicity for combinatorial genetic...cost-effective alternative to commercial ladders which also provides a useful yet low-pressure teaching opportunity...less than half the size of either Cas9 or Cas12a. Despite its small size, CasMINI is comparable in efficacy...
  16. Addgene's Tips for Plasmid Quality Control

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    Blog Post
    ... and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above...sometimes have to design new primers to sequence the important features. You can use primer design software such...such as Primer3 to help you design your custom primers. Remember, when designing a sequencing primer, you... to differ between plasmids, and selecting or designing a primer specifically to target this region, as...instructions on other factors to consider when designing your primers can be found here. Finally, you will... to Verify Your Plasmid Learn How Addgene Uses Barcodes to Track Samples Resources on the Addgene Website...
  17. Plasmids 101: Screening Strategies Used in Plasmid Cloning

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    Blog Post
    ...about colony PCR in this blog post.   Figure 2: The steps of colony PCR.   Sanger sequencing Sanger...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a  screen comes in. Let’s...blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse...Sanger sequencing determines the precise order of nucleotides within a DNA molecule, in this case a plasmid... of the information you need, so you'll want to design a custom primer. Because Sanger sequencing can ...
  18. All in a Twist: dsRNA

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    Blog Post
    ... would release them into the cytosol. Fig. 2: Endogenous sources of duplexed RNA DNA:RNA hybrids...into agriculture as species-specific pesticides and insecticides, an advancement over previous chemical...prone to degradation – those are the words that describe RNA! Double stranded? Well, that’s just for DNA...also known as RNAi, are short, dsRNA molecules designed to degrade mRNA transcripts. RNAi activates the...sequence is found (typically RNA that the RNAi is designed against), RISC can repress translation and/or ... and repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular...
  19. Gendered Innovations: Why Does Sex of the Cell Matter?

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    Blog Post
    ...state-of-the-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on increasing...using both sexes in basic research is excellent design), but they used all female stem cells—this was ...environmental factors. How to include sex in experimental design Recognizing the importance of analyzing sex, the... above, another excellent resource for research design is Kalpit Shah et al., “Do you know the sex of ...Corporations need to integrate these insights into product design. But eyes have been opened—and we cannot return...
Showing: 461 - 480 of 639 results