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...Covalently to DNA via a Phosphotyrosyl Linkage.” Molecular and Cellular Biology 5(11): 3274–79. PubMed PMID: 2427927...blogger Katrin Michel. Cre-lox is an incredibly popular and powerful site specific recombinase (SSR) system... and Sadowski 1985). The FLP system works in a similar fashion to the Cre system. The FLP recombinase ...excision, inversion, and translocation of DNA in a similar fashion as between loxP sites (For details see ...mammalian cells (usually grown at 37 °C). Smart molecular evolution led to the identification of FLPe which...by adding a nuclear localization sequence (SV40 Large T nuclear localization sequence) and expressing ...codon optimization resulted in FLPO, which shows similar efficiency to Cre (Kranz et al. 2010). These differences...

...of neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in the...reliable primary antibodies for their detection, particularly when antibodies to the POI are non-specific,...discussed above, weakly expressed proteins in particular are difficult to track with these classic probes...researchers in Loren Looger’s lab have developed new molecular tags combining the advantages of both (Viswanathan...used to amplify the direct smFP signal within sub-cellular structures. For instance, smFPs have been used... the stratum oriens and the stratum lacunosum moleculare were better resolved with smFP_FLAG than GFP....ability to distinguish weakly expressed and/or similar proteins Through their multiple, highly specific...

...and is particularly excited to hear about novel developments in the fields of gene and cellular therapies...(e.g. a postgraduate degree in law, an MBA, or similar), there are many roles out there that require exactly...to manipulate cells (manufactured by one of the larger biotech companies), and as far as I remember, it...experience. With this valuable training behind you (and largely only that is necessary), there are opportunities...Technical Support Advisor, Technical Correspondent or similar. While there isn’t a clear distinction between ...candidate requirements (and maybe also the proposed salary range) should help. In smaller companies, technical...primarily office based, and email/phone are the most popular ways to communicate with customers. Online live...

...use different RT enzymes and performed large edits with similar or better efficiency than PEmax, the current...Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old...REcipient Cells Through Envelope Design). This modular approach combines an interchangeable set of fusogenic...and in vitro applications.   Figure 2: Modular features of DIRECTED particles where the fusion... et al. (2023). Cell type-specific delivery by modular envelope design. Nature Communications, 14(1), ... TurboID and APEX2, are localized to different cellular compartments. They showed APEX2 can use an orthogonal...sequential labeling by TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled proteins...

... several of our most popular CRISPR pooled libraries and one of our most popular barcoding libraries as...formation titering assay or similar protocol. After functional titer is determined, larger lentiviral preps can...depositing lab. (For our more popular libraries, you might receive a similarly-sized aliquot from an amplification...first step will be to amplify the library. This is similar to amplifying a plasmid: transform E. coli with...place to start is with a protocol for a library of similar size and backbone.  Verification Once your library...representation of your amplified library should be very similar to that of the original library. Like amplification...plasmids in the library.  Spinfection If you have a large number of cells to transduce, you may want to consider...

...fluorophores that have a similar brightness - even though in the most popular pair, CFP has five-fold ...technique has revolutionised the way we apprehend molecular complexes and is still a very useful tool.    ...living cells using a microscope. Historically, molecular interactions were detected by indirect means often...FRET reduced the scale of our perception about molecular interactions. What is FRET? In their JCB 2003...is transferred non-radiatively from an excited molecular fluorophore (the donor) to another fluorophore...fluorophore (the acceptor) by means of intermolecular long-range dipole-dipole coupling." The emission of the acceptor... Constructs to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian...

...Eindhoven has similar interest in FRET-based sensor proteins, particularly for the intracellular imaging of...effort to understand the connection between molecular- and cellular-level events and behavior. It’s a problem... want them to go. Griesbeck is motivated by a particular interest in monitoring the activity and biochemistry..., Griesbeck adds that this color change has a particular advantage for observing events taking place inside...optimized by testing various linker configurations in a large-scale functional screen in bacteria. Sensor variants...quite a robust way to make a FRET sensor with a large dynamic range,” he says. Merkx recently applied ...Hot Plasmid page, where we highlight some of the popular & newly available plasmids at Addgene. If you're...

...salts. Capillary electrophoresis Quantifying DNA using capillary electrophoresis is similar to quantifying...first before larger fragments. Figure 1: The quantification of a RNA ladder using capillary electrophoresis...Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for molecular biology...electrophoresis. Smaller fragments migrate quicker than larger fragments. These fragments are quantified over ...time. Smaller fragments migrate more quickly than larger fragments. Image from OpenWetWare. This method...resources on the Addgene blog Read more about molecular biology protocols and tips Find the best way to...

...unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent problem... Numbers in the large colored circles are rough approximations of the total number of CRISPR plasmids...plasmids for that particular organism available at Addgene. Percentages represent the fraction of that total...function. One huge reason CRISPR has become such a popular genome editing tool is its developers’ willingness...that, for many groups, may not be worthwhile. Particularly if in the end all they want to do is to quickly...of insert) and the reduced titer associated with large inserts can make lentivirus production frustrating...libraries now available in lentiviral format Another popular set of tools in the repository are CRISPR pooled...

...and more. MoClo modular cloning system Synthetic biologists have developed a modular cloning strategy... Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector that can be used to generate random...mechanism for sgRNA delivery and providing a means for large scale functional screens. For more information on...the cloning process for complex DNA constructs, particular for those involving recombination elements such...such as FRT or Lox sites, and provides a modular system for easier exchange and re-use of existing elements...provides improvements to classical cloning methods, particularly for complex transgenesis constructs. Kirchmaier... kinases Receptor tyrosine kinases (RTKs) are a large class of cell-surface receptors which play a critical...