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... Potential Applications in Biotechnology. Critical Reviews in Biotechnology, 24(4), 155–208. https://doi.org...virulence plasmid are now used routinely in biotechnology to transform plant cells. Degradative plasmids...from the benefit of resistance plasmids, which protect the host from attacks by other organisms. To conserve...and others have proposed developing new cloning techniques based on these vectors (Shareck et al., 2004)...Nature’s toolbox for new and exciting molecular technologies!  This post was originally written by Jessica...

...Many, many techniques are available to assess protein-protein interactions. One popular approach is to...under physiological conditions. Shyu et al. Biotechniques. 2006 Chang-Deng Hu Green sfGFP Split ...reassembly is a fast, efficient, and robust method for detecting protein-protein interactions in vivo. Blakeley...under physiological conditions. Shyu et al. Biotechniques. 2006 Chang-Deng Hu Venus Improved N-terminal...a high signal-to-noise ratio. Kodama et al. Biotechniques. 2010 Chang-Deng Hu mVenus pET-BiFC contains... labeling for single-molecule imaging TagBiFC technique allows long-term single-molecule tracking of protein-protein...

...revolution for antibodies. Recombinant antibody technology makes it easy for scientists to share, remix,...Antibodies are used by scientists around the world to detect, purify, quantify, deplete, and visualize proteins...antibodies or monoclonal hybridomas, but those techniques have several drawbacks. Polyclonal antibodies...proposed a shift to recombinant DNA-based antibody technologies (Bradbury & Plückthun, 2015a).   Figure...srep31730 Tsuruta LR, dos ML, Moro AM (2018) Display Technologies for the Selection of Monoclonal Antibodies for... for Clinical Use. In: Antibody Engineering. InTech Additional resources on the Addgene blog Read our...

...about Mammalian shRNA Tools for RNAi! Cre-Lox technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral... and Cre-recombinase. The Cre-lox system is a technology that can be used to induce site-specific recombination...knockout, and selection marker removal. CRISPR technology pdCas9-humanized (Plasmid #44246) Expression ... the pLKO.1 vector can be found here. Cre-Lox technology Puro.Cre empty vector (Plasmid #17408)   This...lentiviral prep from the Addgene viral service. CRISPR technology lentiCRISPR v2 (Plasmid #52961) This is a 3rd...Maetzig T, et al. Gammaretroviral Vectors: Biology, Technology and Application. Viruses. 2011. PubMed PMID: ...

...Interactions Difficult to detect weak and/or transient PPIs. Many PPI identification techniques rely on protein ...version of the proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that...proteins. No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “on/...extended incubation period could make it difficult to detect proteins with short half-lives. Interested in using...Additional Resources on the Addgene Blog Learn about techniques for visualizing protein turnover Learn about ...

...wonder about the origins of some of the common techniques or tools you use in the lab? Take for instance...development of antibodies as tools allowed us to detect and capture proteins of interest to better study...mid-late 20th century with the advent of hybridoma technology and monoclonal antibodies. However, it was (and...this issue by developing generic strategies for detecting cloned genes (Munro and Pelham, 1984). The strategy...be chemically treated before the antibody could detect them (Munro and Pelham, 1984). In their paper they...Munro S, Pelham HR (1984) Use of peptide tagging to detect proteins expressed from cloned genes: deletion ...

...learning techniques to make sense of biology, with a special focus on gene editing techniques. References...Prime editing is a versatile genome editing technology that allows precise modifications of DNA (replacements...features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org/10.1038/s41587-023...and product purity by deep learning. Nature Biotechnology, 41, 1151–1159. https://doi.org/10.1038/s41587...efficiency across diverse chromatin contexts. Nature Biotechnology. https://doi.org/10.1038/s41587-024-02268-2 ...

...localization of your protein of interest is to make it detectable. Visualization can be achieved using fluorescent...image analysis software are highly sensitive to detecting light signals not visible to the human eye and... imaging such as bleed-through (the incorrect detection of one fluorescent signal in a channel setup for...environments. Since colocalization relies upon the detection of 2 independent fluorescent signals and their...practically speaking, for common lab microscopes the detection limit is 200 nm. The size of most proteins is ...is below 10 nm and therefore colocalization detected in microscopy cannot be interpreted as direct molecular...

...These developments in microscopy coupled with techniques that make tissues transparent are enabling microscopes...microscopes to visualize the cellular architecture of whole tissues in 3D with unprecedented detail.  ...absorption.  Finally, the photomultipliers used to detect light in confocal microscopes are less efficient...the sample surrounding the focal plane of the detection lens and thus there is no out of focus light. ... been significant advances in tissue clearing techniques during this century.  The first successful attempt...//www.physikalischebiologie.de http://www.lavisionbiotec.com/ultramicroscope-overview.html http://...

...proteins fused to the halves of the split FP. This technique is also is also called Bimolecular Fluorescence...Therefore, choose FPs that your system can excite, and detect the emission. For example, if your microscope has...have a filter that will pass blue light to the detector/camera, then BFPs are of no use to you. When using...  have a broad range of emission which will be detected by longer-wavelength filters (e.g. GFP also emits...looking for more fluorescence microscopy tools and techniques to aid your work, head over to greenfluorescentblog...2011) “Modern fluorescent proteins and imaging technologies to study gene expression, nuclear localization...

...where you assume your technical skills define you and that the other not-so-technical skills you’ve built... individual with more going for you than your technical expertise in a specific field. While searching...under your belt from your last job. You knock the technical portion of the interview out of the park. It’s...hiring team will also want to examine not just technical skills, but also transferable skills, before offering... make hiring decisions. Though having all the technical skills certainly matters in the job market, it...

...emerging family of "GPS for the genome" techniques Technique What It Locates ChIP-seq Proteins (for...combination with the techniques from Table 1, provides us with powerful GPS-like techniques to explore dark...pairs in length. An emerging family of sequencing techniques function as a kind of “GPS for the genome” to...bound to modified proteins called histones. We can detect modified histones with “GPS for proteins,” ChIP-seq...

...-length viral genome (Figure 2). While it is technically possible to encode all four viral genes in the... (see Uses of Minigenome System). From a more technical standpoint, a plasmid with all four genes and ... gene by altering the amount transfected (see Technical Tips). Minigenome replication and transcription...modeling genome replication. These products can be detected by Northern blot analysis. In addition, the polymerase...viral polymerase. Alternatively, the mRNA can be detected directly by Northern blot analysis or RT-PCR. ...compounds which often target the viral polymerase. Technical tips It is important that all of these plasmids...

...genes that fell into 3 categories based on the architecture of the CRISPR protein in question: C2c1, C2c2...binding a user-defined RNA sequence could be used to detect single molecules of an RNA species with high specificity...tumor mutations within cell-free genomic DNA, and detect COVID-19 (Joung, Ladha et al., 2020). Cas13a in...CRISPR community is an essential part of the rapid technology development seen in the field and as always, ...CRISPR-C2c2 enable guide-RNA processing and RNA detection." Nature 538(7624) (2016):270-273. PubMed PMID...Gootenberg, Jonathan S., et al. "Nucleic acid detection with CRISPR-Cas13a/C2c2." Science (2017): eaam9321...

.... Instead, you can choose a molecular cloning technique that will work well with a given set of resources... invention in the late 1990s, Gateway cloning technology has become very popular as a rapid and highly...and its advantages. An introduction to Gateway technology The Gateway cloning method, developed by Invitrogen...clone in their desired DNA sequences. Gateway technology relies on the two reactions described below: ... more information. How to clone using Gateway technology To better understand the process, we’ll walk ...References 1. Chee JY, Chin CF (2015) Gateway Cloning Technology: Advantages and Drawbacks. Clon Transgen 4:138...

...types of replicate controls: technical and biological. In general, technical replicates can be thought of... more in depth information. Technical replicates An example of a technical control is the transfection...experiments employing gene targeting or genome editing technologies, such as RNAi or CRISPR, non-targeting controls...the independent variable. Replicate controls: technical and biological Reliable results are reproducible...

...social contract citizens enter into in order to protect their rights (2). Less formally, governments are...for a given policy. In exchange, the government protects (and often funds [3]) scientists as well as other...Aeronautics and Space Agency, the Environmental Protection Agency, and the National Institutes of Health...impact (15). Diversity and inclusion in science, technology, engineering and mathematics (STEM) (17). This... 27). In order to achieve the breakthroughs, technologies, and treatments promised by modern research,...Trump’s first budget Environmental Science and Technology: Crossing The Imaginary Line The New York Times...

...affect a protein of interest’s function. This technique relies on the generation of a plasmid library ...pot saturation mutagenesis as a quick and easy technique that can be used to generate complex libraries...throughput.       Gibson Assembly: An easy-to-use technique that joins homologous DNA fragments in a single-tube...mutagenesis The one-pot saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach...primer pair to generate an individual mutant, this technique uses mutagenic primers that tile across a region...protein function on a massive scale." Trends in Biotechnology 29.9 (2011): 435-42. PubMed PMID: 21561674. ...

...genome engineering is done with recombineering, a technique that utilizes phage recombination machinery to...spacer sequence, and these rare events are easily detectable using PCR. The system also functions in S. pneumoniae...MAGE (Multiplex Automated Genome Engineering) technology (Ronda et al., 2016). pMA7CR_2.0 expresses lambda...Scientists have begun to realize the potential CRISPR technology can have on the human microbiome. You can consider...produce sequence-specific antimicrobials. Nat Biotechnol 32:1146–1150 . https://doi.org/10.1038/nbt.3043...efficiently delivered RNA-guided nucleases. Nat Biotechnol 32:1141–1145 . https://doi.org/10.1038/nbt.3011...bacterial genomes using CRISPR-Cas systems. Nat Biotechnol 31:233–239 . https://doi.org/10.1038/nbt.2508...

...address these limitations, labs have developed techniques like CREATE (Deverman et al., 2016), M-CREATE... on reagents and protocols, visit https://clover.caltech.edu. Figure adapted from Challis et al. 2022....deposited by the Gradinaru lab. You can visit our Caltech Systemic Capsids page to learn more about that ...It shows lower expression in the liver and no detectable transduction in the CNS. In the ENS, it has a...need more options) we recommend you check out Caltech’s CLOVER Center’s extensive repository of published... Gradinaru V, Fox AS. (2023). Advances in AAV technology for delivering genetically encoded cargo to the...throughout the non-human primate brain. Nature Nanotechnology, 1–11. https://doi.org/10.1038/s41565-023-01419...