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  1. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...Almo, Institute for Protein Innovation  1:18:45 - 2:01:18 (end) - Panel Alex Tucker (Ginkgo Bioworks)...their curation of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to ... and here I summarize what I learned both in preparation for my talk and at the minisymposium. You can...address this issue directly by creating well characterised antibodies against all human extracellular proteins...stories and read papers wherein a researcher tries 5 separate experiments at once to get confirmatory data for...
  2. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle work in any plant species...
  3. Let There Be LITE Plasmids

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    Blog Post
    ...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...ingredients - CRY2 and CIB1 – which they borrowed from Arabidopsis thaliana. The results are light-inducible transcriptional...
  4. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...single cell level), are separated by, for example, cell sorting. The separated cells are subsequently ...which gRNAs are differentially expressed in the separate groups and, as a consequence, which genes are ...the protein following stimulation would then be separated by sorting, and subsequently sequenced to identify...suggested gRNA sequence including adapters. Output_UserInputParams: Contains all the information that you ...al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach." Nature methods...
  5. Finding Your Science Policy Path

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    Blog Post
    ...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...the work you do to creatively connect seemingly disparate individuals and ideas toward solving one problem...evidence-based university policy to combat sexual harassment in academia or a large company who decides how...
  6. Antibodies 101: Validation

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    Blog Post
    ...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...antigen - Recognition of an immunogen does not guarantee recognition of the target antigen. You want to... transfect it into your favorite cell line. In parallel, you transfect cells with another plasmid not ...
  7. RUBY-Red Siliques

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    Blog Post
    ...make it easier to study these mutants.   Fig. 2: (A) Betacyanin is produced from tyrosine by the activity...occurs in T0 plants (from callus) or T1 plants (Arabidopsis floral dip). In the next generation the CRISPR...planning to use it to simplify my work with an Arabidopsis mutant that can’t set seeds. I study the development...   Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following floral dip transformation next...without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing seeds from ... the Plant Genome Using CRISPR/Cas9 Tips for Arabidopsis Transformation CRISPR 101: CRISPR-Mediated Plant...
  8. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

    Type
    Blog Post
    ... stabilization of unknown bacteria. Figure 2: Christina Mork collecting samples at the hot spring...microfluidic parallelization to sequence this highly complex hot spring sample. The sample was separated into ...Quake Lab member joined for half of the trip. Preparations ahead of time included contacting YNP staff ... Paul and Anastasia (Potanina) Neddersen characterized a few samples by 16S rRNA gene sequencing which...thermostable Cas9 enzymes have previously been characterized: GeoCas9 from the Doudna lab and ThermoCas9 ...
  9. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ... ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...fluorescent labels. In Nature Methods (Vol. 19, Issue 2, pp. 149–158). Nature Research. https://doi.org/10.1038...help. Here, we will outline some fundamental characteristics to consider when selecting a dye or ligand ...Fluorescence microscopy takes advantage of the characteristics of these fluorophores to enable highly specific...receptor conformation and signaling activity​​ (Sridharan et al., 2014). It's also crucial to evaluate how...impacted by both the excitation coefficient, characterizing the strength a species absorbs or reflects ...considerations regarding the choice of fluorescent dyes (Sridharan et al., 2014). Manufacturers will typically recommend...
  10. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2 coat... 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. "The green fluorescent protein." ...microscopy (University of Amsterdam). He develops, characterizes, and uses genetically encoded fluorescent probes...
  11. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ...containing complementary polynucleotide chains. J Virol 9(2): 394-396. Lecomte E, Tournaire B, Cogné B, Dupont...facilities producing a large number of viral preparations in parallel, there is a very real risk of mixing up...Léger A (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded Virus...confirm the identity and serotype of our AAV preparations. We recently published a description of the ... For labs that produce a small number of AAV preparations this level of quality control is sufficient ...AAVs and the TA-based ligation and tagmentation preparation approaches described previously require a preliminary...effective substrates for tagmentation-based library preparation procedures. This finding eliminates the double-stranding...
  12. Editor's Choice, September 2016

    Type
    Blog Post
    ... of my favorite molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique...That’s it for now. As I indicated in the opening paragraph, I’m currently on my way back from a very successful...
  13. Hot Plasmids and Viral Preps - July 2021

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    Blog Post
    ...method relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity...are also known to trigger seed germination of parasitic plants, this approach was also used to build and...ratiometric fluorescent biosensor based on the parasitic Striga hermonthica SL receptor HTL7. This novel...compounds that have the potential to regulate the parasitization of cereal crops. As a result, agricultural ...
  14. Addgene @ Keystone: Thoughts on Precision Genome Engineering and Synbio

    Type
    Blog Post
    ...in just 2 years and excitement about how much more there is left to do with the characterization and implementation...relatively isolated resorts creates a sense of camaraderie with fellow attendees from the moment you arrive...motto for scientists who contact us and want a guarantee that their experiments will work (yes, this happens...
  15. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...interferon gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of...thanks to our guest blogger Subhadra Jayaraman! Subhadra Jayaraman is currently a doctoral candidate at...post was contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University... level of progression, anatomical location, characteristic (solid localized tumors or systemic tumors)...
  16. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...establishing the dynamic range of your FRET assay (Figure 2). These labeling methods are freely interchangeable...PMID: 20161833. PubMed Central PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding...), quantifying antibody/antigen interactions (Saraheimo et al., 2013), and visualizing structural changes...fluorophores, it’s best to consider these cases separately when talking about the tools that are available...tools for labeling reagents and structurally characterizing large ion channels involved in skeletal muscle...40.3 (2006): 279-286. PubMed PMID: 16908183. 10. Saraheimo, Satu, et al. "Time-Resolved FRET-Based Approach...
  17. The Power Behind NGS Plasmid Validation: seqWell

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    Blog Post
    ...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...many labs. However, the high costs of sample preparation for high-throughput next generation (NGS) sequencing...Addgene to develop and apply our plexWell™ Library Preparation Technology for NGS-based sequencing and confirmation...of barcoded fragments is known as a *library*. Parallel sequencing of the fragments within a library allows...understated problem in multiplex NGS library preparation. The challenge here is that, as more samples ...minimize these issues, researchers usually include a separate normalization step where individual input DNA ...
  18. Easing the Protein Purification Process with pCri

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    Blog Post
    ...PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary T. Britton, and Anne S...number of disulfide bonds in your protein - two parameters that affect how well E. coli can produce your...
  19. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    Blog Post
    ...motion videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time...gotten easier now that I identify with the main character in the video. We’ve grown attached to the work...“WOW, now I have my own laboratory” (the main character is much more than a piece of plastic for us now...
  20. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...access this tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence...complement sequence.  Filter DNA tool: Removes non-DNA characters, such as numbers and blank spaces, from the text... primer dimer estimation, and other primer characteristics. NEB Data-optimized Assembly Design (DAD): ...
Showing: 321 - 340 of 486 results