...results.  Replicates Replicates are another critical control in biological experiments. Replicates can be...inclusion of biological replicates such as multiple mice in each group and C, technical replicates or repeated...meaningful results.  Biological replicates are distinct samples that are treated identically, used to assess...recorded would be due to natural biological variation. This information is critical for correctly interpreting...be technical or biological and are included to improve the precision and accuracy and therefore the statistical...statistical power of an experiment. Technical replicates are repeated measurements of the same sample ... rely on the results of a single mouse. Biological replicates help you to determine which responses arise...

... Standard Curve?  Amplification Efficacy Calculation Amplification Efficacy Assumptions Standard...increase your understanding and technical proficiency with common applications - now let’s dive right in! ...differences of twofold or greater with good statistical significance (Taylor, 2017). With the right conditions...single peak, indicating one amplicon and no primer dimers. (b) A shoulder peak indicating primer dimers...values within each set of triplicates to understand variation due to technical error.  Plate readouts can... be less than 0.1.  If you suspect that amplification efficacy differences may be a concern in your reaction...genes can have different efficacies DART-PCR No Raw Data Statistically defined log-linear phase...

...Origin of Replication? See our Plasmids 101: Origin of Replication blog).  The replication problem Plasmids...directly inhibit replication by binding to the origin of replication and blocking replication machinery (del... of microbial plasmids: classification and identification based on replication and transfer systems and...plasmid that contains the origin of replication and the replication control machinery (Need a refresher... that inhibits replication when copy number in the cell is high, but allows replication when copy number...either indirectly inhibit replication through inhibiting translation of replication machinery proteins, or...antisense RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated...

...two types of replicate controls: technical and biological. In general, technical replicates can be thought...of the independent variable. Replicate controls: technical and biological Reliable results are reproducible...independent and are typically derived from one source. Conversely, biological replicates ARE independent ... more in depth information. Technical replicates An example of a technical control is the transfection...independent variable. Biological replicates In any given set of experiments, the biological controls are more...results. Typically, plasmid-based experiments employ transfection, negative, positive, and replicate controls...thought of as "reproducibility controls". Biological replicates are what makes up your sample size (aka ...

...non-bench research career such as law, medical/technical writing, clinical, regulatory, product development,...specialty occupation" as requiring theoretical and practical application of a body of highly specialized ...Further, this post may contain technical inaccuracies or typographical errors.  No action should be taken...18, 2017, President Trump signed a "Buy American, Hire American" Executive Order which sets broad policy... industry such as technology transfer, communications, medical writing, compliance, core facility management...received 116,224 employment based green card applications and approved 91,023 during the first 3 quarters...options discussed in this post benefit from the applicant thinking about the process early, keeping good...

...as synthesis modifications when purchasing your gRNA. Pro tip: modifications are typically incorporated...switches. Stabilizing gRNA modifications  Sugar phosphate backbone modifications The recognition sequence...phosphorothioates with 2’-O-methyl modifications, proved more stability than just one modification alone (Hendel, et al...modified) for optimal stability and efficacy.  Base modifications What is more stable than RNA? DNA! ...stabilizing modifications listed above were developed before CRISPR-Cas9, for use in applications like siRNA...oligonucleotides. Some of these synthetic modifications, specifically ones which do not contain a 5’ triphosphate...your next CRISPR experiment.    Chemical RNA modifications to improve stability.   On-off switches...

...in science communication or policy, being able to communicate your work to a non-technical audience is...knowledge and applications generated from this research. At both a practical and ethical level, scientists...benefits as well as the personal, social and ethical implications of genetics. In these conversations, pgEd...training beyond the biological sciences, as well as to acquire leadership and communication experience. These...science policy and science communication A career in science communication or policy requires broad interest... in our field of interest. We often communicate in very technical and jargon-filled language in academic.... Also seek out opportunities to communicate science to non-technical audiences. Many universities or ...

...illumination on biochemical processes that are crucial in the world of biological research, medical therapy, ...components, typically a protein of interest. Fluorescent dyes and ligands vary in their chemical structure...the affinity tag they typically bind to. Stability of fluorescence: For applications that involve ranges...recruitment to receptors, indicating their agonistic nature. The potency and efficacy of these ligands can ...their product descriptions, or technical manuals. Specific applications and matching dye/ligand choices... in biological research. This does make finding the right dye for your experiment more complicated. We...ligands with G protein coupled receptors. In Biochimica et Biophysica Acta - Biomembranes (Vol. 1838, Issue ...

... norvegicus genes! Mexican axolotl (Ambystoma mexicanum) Ambystoma mexicanum, aka the axolotl. Image...essential aspect of biological research that has defined our understanding of biological processes and developments...studying genetic modification for their ease of manipulation and their ability to be replicated. Because E....providing insight into some of the most significant and fundamental biological processes. Many of their genetics...tools in developmental and biomedical research.  Xenopus, aka African frog. Image from Holger Krisp...Advances on Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306...Organism. News-Medical. Retrieved on September 18, 2020 from https://www.news-medical.net/life-sciences...

...the grant application process. When my PhD advisor first asked me to write a grant application due the ...pursue. Proposal writing: Improves technical writing and communication skills, Elucidates the big picture...information required for a successful grant application. It typically includes overview information, key dates...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...Research at universities typically requires funding from a variety of government and academic institutions...advisor alone applies for these competitive grant applications; however, in some cases, your advisor may choose...

...Other than a theoretical maximum value, physical titers don’t necessarily give any indication of the infectious... red) report physical titer, which is measured by qPCR.   Physical titer is typically calculated by... you? Physical titer: the concentration of viral particles containing viral genomes. Physical titers ...qPCR or other DNA quantification methods - see below), since each viral particle typically contains one viral...facilities report the physical titer of a viral solution (Figure 2). Because physical titers are used for...titer values. Finally, because physical titer does not give an indication of infectious titer, consider...that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type...

... of the American Chemical Society 123.48 (2001): 12123-12125. Journal of the American Chemical Society...discovery–a review of theoretical aspects and recent applications." Current Chemical Genomics and Translational...originally developed to treat heavy metal intoxication in chemical warfare [Vilensky & Redman, 2003]). Most...al. "Designing logical codon reassignment–Expanding the chemistry in biology." Chemical Science 6.1 (2015... of a biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal of the American Chemical...laboratory, then FRET experiments are a viable biophysical path to answers. What is FRET and what's it...Fluorescence resonance energy transfer (FRET) is a biophysical interaction between two fluorophores. When the...

...underlying all communications efforts.  Scicomm definitions (as used by Addgene)    Science communication, or science...time engaging with science communication efforts.  Scientific communication or scientific writing is peer...to note that it is the actual communication itself, not the communication method, that determines if something...scientific or science communication. A Tweet is still considered scientific communication if the language is...Series, where we’ll be talking about science communication, best practices in the field, and even getting...getting into some active science communication research.   This summer, we’re making time to talk about ...about my favorite thing - science communication (scicomm)! We have so many fun topics planned for this summer...

...epigenome (the sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting...DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic...Preps for Epigenetic Modification Why use epigenetic modifiers? Epigenetic modification is certainly not ...Nature Communications 8: . https://doi.org/10.1038/ncomms16026 Find plasmids from this publication at Addgene..., 2017 and updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...mechanisms remain unclear. Today, targeted epigenetic modification at specific genomic loci is possible using CRISPR...

...or DYKDDDK 1 Affinity and Purification Good for antibody-based purification; has inherent enterokinase...site GST Large Protein 26 Purification and Stability Good for purification with glutathione; protects...used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity and Purification Very small...GKPIPNPLLGLDST   1.4 Affinity and Purification Good for antibody-based purification Combo and cleavage tags...increasingly common. Tandem affinity purification (TAP) once referred specifically to a combo tag comprised of ...multitude of uses including (but not limited to) purification, detection, solubilization, localization, or...overexpress and purify proteins can face any number of technical challenges depending on their protein of interest...

...differing in pigmentation." Biochemical and biophysical research communications 78.1 (1977): 237-243. PubMed...one that can be achieved using pharmacological or electrophysiological methods (Deisseroth, 2011; Mei ...recombination. Purification and properties of the Cre recombinase protein." Journal of Biological Chemistry...Edward S., et al. "Millisecond-timescale, genetically targeted optical control of neural activity." Nature ..., et al. "Next-generation optical technologies for illuminating genetically targeted brain circuits." ...and Jinfang Zhu. "Immunologic applications of conditional gene modification technology in the mouse." Current... R. Thompson. "Overview on research and clinical applications of optogenetics." Current protocols in pharmacology...

... construct for every application. Plus, the photophysical properties of chemical dyes provide many advantages...Few Complicated Non-natural amino acids 1 Various Many Complicated   Applications Organic... W. (2011). Chemical Tags for Labeling Proteins Inside Living Cells. Accounts of Chemical Research, 44...If you take a look in a typical lab freezer, you’ll probably find lots of different plasmids containing...new green fluorescent protein, some affinity purification tags, another version with that same red fluorescent...substrate. If the substrate was linked to, say, a chemical dye or other functional ligand, your protein of...advantages over fluorescent proteins, opening up new applications.   Figure 1: Live-cell labeling strategies...

...Spatially Confined Persistent Diradical Emitter. Journal of the American Chemical Society, 145(25), 13615–13622... tools for familiar applications. But how does one invent a tool for applications that don’t even exist...organisms) are opaque, which makes most biomedical applications of these tools difficult or impossible....of developing new microscopy tools for biological applications across a wide range of topics. He embraces...., Timmel, C. R., & Hore, P. J. (2016). Chemical amplification of magnetic field effects relevant to avian...phenomenon, improve the tool, and develop new applications for magnetoresponsive proteins. Motivated by...tool with magnets would enable new research and applications that don’t yet exist, but it wasn’t clear that...

...of Various Motifs is Critical for Initiation of Replication. Journal of Biological Chemistry, 273(14), ...to utilize diverse host replication machinery. For instance, the ori typically contains iterons that serve...ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various hosts...the ori and allow for replication. However, if plasmids have their own replication machinery such as Rep...different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow... while others have a broad host range and can replicate and stably maintain the genes they carry among.... These features are: The plasmid origin of replication (ori) contains structural elements that are versatile...

... your own research! Classification Structure Principles of CRISPR classification While CRISPR systems... CRISPR-Cas systems. Philosophical Transactions of the Royal Society B Biological Sciences, 374(1772),...Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied...properties. This led to a need to develop a classification system for the array of CRISPR technologies...technologies. Learning the general principles of this classification system will help you gain a deeper appreciation...multi-pronged analysis to sort effectively. The current classification system relies on a combination of sequence ...a similar function. After analysis, CRISPR classifications were placed into a fairly simple hierarchy:...