We narrowed to 1,052 results for: FER
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TypeBlog Post...widely utilized across many different cell types and each one requires different elements for vector propagation... features of yeast vectors and explain how they differ from the bacterial elements previously described...auxotrophic markers and includes the associated references. Yeast selection marker What does the marker...
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Kiran Musunuru on the Newest TALEN Genome-Editing System
TypeBlog Post...stem cells edited to carry mutant versions of 15 different disease-associated genes. Addgene spoke to Musunuru...mutations. Once that's done in stem cells, we can differentiate those mutant and wild type stem cells into any...all have the mutations but they will also have different genetic backgrounds. It becomes more of an apples-to-oranges...suggest TALENs may be cleaner than CRISPR/Cas9. Reference: Quirong Ding et al. "A TALEN genome-editing system... -
Addgene: The First Twenty Years
TypeBlog Post...organizations, knocking on doors, and attending conferences. And when CRISPR arrived in 2012, our repository...Part of that work meant facilitating the Material Transfer Agreement process, which was a major barrier for...2013, which continue to grow today. “I generally refer people to Addgene for the basics of molecular biology...’s booth at the Society for Neuroscience 2023 conference. In 2020, the world changed in response to the... -
Antibodies 101: Fab Fragments
TypeBlog Post... for your specific scenario. References and resources References Bates A, Power CA (2019) David vs...is relatively easy to physically divide up the different regions, both through genetic and proteolytic ...broken up, there are a few commonly available different fragments that retain antigen binding (Figure ... -
All in a Twist: dsRNA
TypeBlog Post... disease, and much more! Resources and References References Wang, I. X., Grunseich, C., Fox, J., et ... dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with bidirectional...blog CRISPR 101: RNA Editing with Cas13 RNA Interference in Plant Biology ... -
RNA Extraction Without A Kit
TypeBlog Post...final RNA sample(s) are resuspended in water or TE buffer, store them in a -80°C freezer to prevent RNA degradation...ultracentrifuge. Figure 1: An outline of the different steps in RNA extraction. Researchers at the... Reagent®, RNAzol®, QIAzol® and sold by many different companies). TRIzol® is an all-in-one acid-guanidinium-phenol... ensure DNA contamination is not an issue. References Allewell NM, Sama A (1974) The... -
Plasmid Tools for Microbiome Studies
TypeBlog Post... microbial species. These also contain the RP4 transfer origin as well as a selectable marker and a desired...and Gammaproteobacteria species isolated from different bee species. BTK can be used to introduce fluorescent...distributed cell division counting plasmids here References Leonard, Sean P., et al. "Genetic engineering... Jennifer Tsang... -
It’s Bioelectric! An Exciting Interdisciplinary Field of Research
TypeBlog Post...negative) the action potential of a cell, the more differentiated and quiescent it is, and the more depolarized... toggled between open and closed states using different wavelength light pulses. Other resources from...bioelectricity, bioelectric researchers in many different fields came together to found the journal Bioelectricity...regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use of genetically... -
The Future of Research Symposium Boston 2015
TypeBlog Post...of trainees? Are there too few or too many at different levels? What does an ideal and productive workforce...a young scientist’s perspective, and the many different paths that biomedical graduates take in their ...policy. Follow David on Twitter @driglar. References 1. Alberts, Bruce, et al. "Rescuing US biomedical...Henry, and Michael Roach. "Science PhD career preferences: levels, changes, and advisor encouragement."... -
Plasmids 101: Golden Gate Cloning
TypeBlog Post...cloning into a modular cloning strategy. Sometimes referred to as MoClo, this strategy uses the Type IIS restriction...256 distinct flanking sequences, sequences that differ by only one base may result in unintended ligation...constructs, assemble standard plasmids parts in different combinations, or other new and exciting applications...constructs in a single, high-efficiency step. References: Sequential amplification of cloned DNA as tandem... -
Making CRISPR Plasmids Using Fragmid
TypeBlog Post...CRISPR experiment. Happy cloning! References and Resources References McGee, A. V., Liu, Y. V., Griffith...chosen because of their broad applicability in different model systems. What’s the error rate? The Doench... pRDA_722 (CMV-driven; lentiCRISPRv2-based; preferred over 512) Mammalian Lentivirus pRDA_789... -
Rabies and Neuronal Tracing
TypeBlog Post...more inputs than was previously thought. The differences in the inputs to each of the two populations ...populations could be used to explain the differences in the functions of these two neuronal populations. Overall...Overall, this study laid the foundation for how different anatomical regions of the brain receive inputs and...complex questions of how the brain operates. References 1. Callaway EM. Transneuronal Circuit Tracing... -
Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools
TypeBlog Post... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover...changes in TagRFP-T fluorescence intensity at different time points, the authors tracked protein activity...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy... FLIM-FRET acceptors here. See Addgene's FRET reference page for many other common FRET vectors. Murakoshi... -
Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog Post...28032891. This is a different version of Split-BioID. BirA* is fragmented at a different site than in Schopp...of the BirA* fusion matters! Schopp et al saw different levels of fusion protein expression and biotinylation...research? Plasmids are available from Addgene! References Schopp, I.M., Ramirez, C.C., Debeljak, J., Kreibich... -
Hot Plasmids February 2024
TypeBlog Post...StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific AAV delivery...quickly saw wide interest, its dimeric nature can interfere with some applications. Several labs have been... viral vectors for functional intravenous gene transfer throughout the non-human primate brain. Nature...the Gradinaru lab, AAV1-X1, was developed by transferring the 7-mer peptide region from variable region... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog Post...more precise editing in the lab. References and resources References Dong D, Guo M, Wang S, Zhu Y, Wang... May 23, 2017 and last updated Jul 23, 2020 by Jennifer Tsang. CRISPR-Cas technology is constantly evolving...loadingExample: AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas... -
Split Fluorescent Proteins for Studying Protein-Protein Interactions
TypeBlog Post...compatible with multiple FP(11) fragments, even from a different base FP (for example, EBFP2(1-10) can be used ... useful. Good luck, and happy experimenting! References and Resources Additional resources on the Addgene....org Fluorescent Biosensors Collection Split Luciferase systems, including NanoLuc®, DULIP, LuTHy Other...inducible genome editing (Feng Zhang Lab 2015) Reference Feng, S., Sekine, S., Pessino, V. et al. Improved... -
Engaging with science and society at pgEd
TypeBlog Post...and epigenetics. So, I sought out a number of different opportunities to broaden my training beyond the...major part of our everyday work is to liaise with different organizations and individuals, to identify ways...collaborating, and to organize events that bring different voices together to talk about genetics. In addition... such as scientific, educational or industry conferences, as well as schools, community events, churches... -
Plasmids 101: FLEx Vectors
TypeBlog Post...white arrowheads). It is necessary to use two different pairs of target sites for this strategy to work...rescued expression of GeneR. Sometimes also referred to as DiO (Double-floxed inverse Orientation),...flip it to "off"). These Cre-off switches may be referred to as DO (Double-floxed Orientation) and could...the genOway press release for more information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling... -
Experimenting in an Open Source lab: from CRISPR to Cats
TypeBlog Post...academic labs, we have the capability to study many different mutants because the Caenorhabditis Genetics Center...domesticated housecat, and seeing how that makeup differs from that of wild cats with the goal of optimizing... space is the Hackerspace website. Markerspaces differ depending on the interests of their community, ...makerspace.ca, Victoria’s first hackerspace which now offers members and the public cheap access to personal...