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...co-factors, thus allowing for broad applications in a variety of experiments. LoxP (locus of X(cross)-over in...have been constructed that express Cre under a variety of ubiquitous and regulated promoters, and many... "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence. Among these are loxN...lox2272 (GgATACtT), and lox511 (GtATACAT). These variant lox sites undergo recombination with other sites...not cross-compatible. Using different lox site variants allows Cre to catalyze more than one specific ...

...COVID-19 research in other labs. Spike protein, variants, and pseudotyping The SARS-CoV-2 virus depends...cellular receptor. To more easily study the spike variants, scientists have developed ways to pseudotype ... has had many opportunities to evolve. Several variants are now circulating and plasmids containing these...pseudovirus and those encoding different spike protein variants that can be incorporated into pseudoviruses. The...at neutralizing the different SARS-CoV-2 spike variants. You can learn more about their research in this...

...modified ABE7.10 accordingly which resulted in variants named “ABE8.x”: “8” stands for the eighth round...x-d.  This resulted altogether in 40 new ABE8 variants. When comparing on-target DNA editing efficiency...replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG...PAM: NNGRRT) to create Sa-ABE8.x-m/d. With these variants, ABE8 could target many additional locations in... base editor: ABE8e Read about xCas9, a CRISPR variant with PAM flexibility Resources on Addgene.org ...

...director of new initiatives at pgEd, I take on a variety of projects in the space of science education, ...genetics pgEd, based at the Department of Genetics of Harvard Medical School, has a mission of raising awareness...ethnic or personal identity. We do this through a variety of activities, including creating educational materials... disciplines, and everyone on staff takes on a variety of projects. With our focus on engaging the community... materials or speak about our experiences in a variety of other settings, including “conventional” academic...

...discussion on effective advocacy strategies on various levels, with panelists including: Kate Stoll, ...Project Lisa Coleman, PhD - Chief Diversity Officer Harvard University  The symposium will culminate with ...appropriate audience and communicating that message in various ways. Workshops will be run by experts in their...Cameron, PhD - Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications...David T. Riglar. David T Riglar is a postdoc at Harvard Medical School. In the lab, he uses synthetic biology...

...in a variety of applications including gene expression profiling, studying copy number variation, and ...nucleotide concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify...concentration Gene expression profiling Copy number variations Quantify RNA levels Molecular Diagnostics NGS...of starting material AAV Titrations Allelic Variations Quantifying low abundance targets Monitoring ...

...downstream applications. As individual libraries can vary significantly, it’s important that we have an amplification...unique plasmids does your library contain?), the variance of size of plasmids in your library (Are the inserts... cells you need total and per transformation may vary depending on the particular type of cells you are...plasmids to overtake larger plasmids, if the library varies significantly in plasmid size). On plates, each... Library Type Name Number of Plasmids Variable Region Size DNA per Amplification Amplification...

... Throughput Low Low Low Efficiency Variable Variable Variable Difficulty Hard Hard Hard These ...system of choice. The table below summarizes the various components included with each expression system...tunable expression of Cas9 and/or gRNA in a wide variety of mammalian cell lines. Useful for difficult to...

...the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem many...George Church’s lab). Addgene has also cataloged a variety of useful molecular cloning protocols, including...comments. Currently you can find protocols from NEB, various academic labs, and now Addgene. I just began uploading...currently boasts over 2,500 protocols covering a variety of bio and biomedical topics. Protocol Online Website...resources, such as protocols, Plasmids 101 articles on various plasmid features, and reference tables (i.e. antibiotic...

...the genetic elements may differ (e.g., a set of various mammalian promoters), the same cloning procedure..., titrate inducible transgene expression, or a variety of other clever applications. In one example, ...downstream applications. The tradeoffs between various cloning methods are listed in Table 2. For example...representation. The specific restriction enzymes used in various chaining-based cloning methods are listed in Table...MXS-chaining method have used the technique to engineer various constructs with applications in flow cytometry ...

...immunoglobulins, the strength of the interaction varies between antibody host species and isotypes. Consequently...factors to consider. Agarose beads are sponge-like and vary in shape and structure. Their porous surface provides...immobilized. The ideal antibody concentration to use will vary between capture antibodies and can be determined...target protein can be eluted. The elution method will vary depending on the specific target protein:capture... of interest on the C- or N-terminus. In this variation, called a pull-down assay, an antibody against...

...at all in E. coli (you can find a variety of databases showing various organisms’ codon preferences online...have different codon usage biases? The reasons for varied codon preferences among organisms aren’t completely...different levels under different conditions. With varied codon usage, an organism can change which proteins...the commercial Rosetta E. coli strains express a variety of tRNAs that are normally found at low abundance...

...either transcript variant led to robust and specific upregulation of the targeted variants without inducing...activation? To summarize, CRISPRa uses a nuclease-dead variant of Cas9 (dCas9) and engineered combinations of ...inducing expression of the non-targeted variant. Moreover, isoform-specific expression of Bdnf resulted in... between the Bdnf I and Bdnf IV overexpression variants were other immediate-early genes associated with...

...Mycoplasma contamination has been shown to arise from a variety of sources such as serum, other cell lines, or ...freeze-thaw cycles on lentiviral stability depends on a variety of factors such as the transgene expressed, the..., this study found that transduction efficiency varied not only between serum from different manufacturers...produce lentivirus may want to consider testing a variety of cation and serum sources to ensure the best ...applications, stick with them! Changes in such variables as cell density, volume, incubation time, and ...

...by Jacob Lazarus, a postdoctoral researcher at Harvard. There’s an astounding number of ways to create...there is also a large amount of user-to-user variability, and in some cases, counter-selection escape ...thanks to our guest blogger Jacob Lazarus from Harvard. Jake received his MD and a PhD in Cell and Molecular...post-doctoral research in the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange ... PubMed Central PMCID: PMC3748948.  Khetrapal, Varnica, et al. "A set of powerful negative selection systems...

...transpeptidase sortase A variant (mgSrtA), which is capable of promiscuously labeling various cell surface proteins...suitable for organoid or in vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the...a small peptide (LPETG). In EXCELL, the mgSrtA variant can be displayed on the surface of a cell of interest...Central PMCID: PMC6126969. Che, Yonglu, and Paul A. Khavari. "Research Techniques Made Simple: Emerging Methods...

... through the gel. For better protein separation, vary the acrylamide content of the gel. For small proteins...assessed from a single western.  There are several variations of the protein transfer step including wet, semi-dry...options targeting your protein of interest from a variety of vendors and you may even find plasmids encoding...Some vendors will also test their antibody in a variety of tissues to demonstrate that the protein staining...unintended cross-reactive proteins. Similarly, if you vary the experimental conditions from those used by the...

...across runs. If the results vary considerably, then the high degree of variability in the process could make...response recorded would be due to natural biological variation. This information is critical for correctly interpreting...of the same sample and are included to assess variability in the experimental protocol. In the experiment...assess naturally occurring random biological variability. If one were studying the effects of tau kinase...

...Zafira Castano Corsino, a junior faculty member at Harvard Medical School/Brigham and Women’s Hospital (BWH...University Miguel de Cervantes, the University of Navarra, and the University of the Basque Country. Mentees...some of the top Universities in USA and Europe (Harvard University, Massachusetts Institute of Technology... Zafira Castaño Corsino is a Junior Faculty at Harvard Medical School/BWH, co-Director of the Mentor Circle...Fernandez-Godino is a Postdoctoral Researcher at MEEI-Harvard Medical School, where she studies age-related macular...

...allow you to optimize the concentrations of your various blot reagents. There are two types of gradients...antibodies, especially polyclonal antibodies, can vary lot-to-lot, this range needs to be empirically determined...antibodies, which have much lower lot-to-lot variability. If you’re running an already-developed assay...of protein in your samples. If you have a wide variety of protein amounts on the same blot, you may want...Add sonication step to protein lysis Splice variants Check literature Posttranslational modifications...use a less sensitive exposure kit No signal variation in highly expressed proteins Antibody dilutions...

...Ligases Small, varies Various Few Complicated Non-natural amino acids 1 Various Many Complicated...HaloTag is actually HaloTag7, and recent specialized variants run up to HaloTag11. HaloTag is now the most popular...any cloning at all! As mentioned above, several variants of these tags have been developed over the years...

... Amy Gilson from Science in the News (SITN) at Harvard University. There are tons of ways you can get ...podcast, the SITN team brings in scientists from a variety of fields as well as students from the ed school...Amy Gilson. Vini Mani is a graduate student at Harvard University, former Co-director of Science in the...     Amy Gilson recently received her PhD from Harvard University, is a former Co-director of Science ...

... sensor. This sensor was engineered from a GFP variant and a DA receptor such that GFP fluoresces when...when the receptor is bound by DA. Two variants of the sensor, dLight1.1 and dLight1.2, exhibit high (submicromolar...GRABDA sensor based on a similar strategy: a GFP variant bound to a DA receptor, such that the GFP fluoresces...detection and measurement of neurotransmitters in various species and while exhibiting complex behaviors....

...my PhD in Biological and Biomedical Sciences at Harvard University and entered a new role here at Addgene... to people. When I started my graduate work at Harvard, I recognized that I would ultimately want to go...the Science in the News Blog, a blog made by the Harvard graduate student group, Science in the News. While... keep content coming also keeps me on top of a variety of fields thereby broadening my knowledge. In addition...

...Interest (YGOI) in a controlled environment under various conditions. The first thing you'll need to decide...protein can be released from the resin and used for various binding or folding assays. Yeast vectors There ...large collection of yeast expression vectors with various markers and promoters. If you need to visualize... downstream effects.  And many more! We have a variety of other types of vectors for most applications...

...small region. This makes them useful for a wide variety of high-throughput experiments. There are currently... to observe clonal dynamics and cell lineage. Variations of this library allow tracking of transcription...’s also highlight a prokaryotic library: the Cultivarium MACKEREL Library, whose acronym means Modular...libraries have been used to systematically explore variations in the spike protein, teaching us more about ...

...impractical to catalog every variant of a feature. Instead, we identified common variants, and then crafted a ...threshold. This issue is addressed by adding more variants to the database. Another limitation is that by...augment the database with new fluorescent protein variants. The future of DNA feature identification What...

...should they decide to produce virus on their own.Harvard researcher Connie Cepko has been helping us with...viral preps are available now? There are a wide variety of tools already available from Addgene as ready-to-use...system. The first two can serve as controls in a variety of experiments, while rtTA can be used to activate...beginning our AAV viral services with these tools. A variety of chemogenetic tools from Bryan Roth’s lab are...

...typically have a wide range of input quantities. This variation is problematic for conventional NGS library prep...range (1ng – 100ng) are normalized to two-fold variation above and below mean read count per sample, and...and the amount of information per read does not vary by DNA input amount. plexWell™ + Addgene = high quality...Boston University, and did postdoctoral work at Harvard Medical School and the University of Toronto. He...

...immune reaction to the treatment. These responses vary depending on the person. However, in silico and ... antigen, pre-existing conditions, etc. that can vary widely between recipients.   In vitro prediction...of the human lymphatic system. PBMCs consist of various immune cells, and can be used as a whole or can...microscopic in-vitro models can be built that contain various components of the immune system. Figure...