...intended application - Remember, just because your antibody performs as expected in one application does ...strategies for antibody validation in different applications. Here, I want to highlight the general suggestions...intentionally generalizable and not specific to certain applications or antibody targets. That said, not every strategy...will be suitable for every antibody or every application. The goal is to show that your antibody has been...strategy to use will depend on your intended application and available resources.   Figure 1: Table...confident in your data. This is a good option for applications like immunohistochemistry on human tissues, ...alternative translation start sites or alternative splicing that could lead to some form of the target protein...

...for both transient and stable gene expression applications in plants. This smaller, easily customizable...Agrobacterium are termed binary vectors because they must replicate in two species: Agrobacterium and E. coli. Like.... coli. However, they also need to be able to replicate in Agrobacterium to ensure that they are maintained... Thus, binary vectors contain two origins of replication, one each for E. coli and Agrobacterium, along...bases each. By switching the E. coli origin of replication and codon-optimizing the Agrobacterium ORI, they...constitutively active M24 promoter has been modified with duplicated enhancer domains and characterized in studies...these properties of pSiM24 are suitable for many applications, the plasmid can also be customized due to its...

...for your application. Image reused form Doman and Pandey et al., 2023 under CC-BY 4.0 License. Find... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...Strebinger et al., 2023, reused under CC-BY 4.0 License. Importantly, even commercial antibodies can...approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting...alkyne-phenol, to label proteins with an alkyne group for click chemistry reactions instead of biotin — a valuable...from Lin et al., 2023 bioRxiv, under CC-BY-NC 4.0 license. This system provides a unique new way to record...

...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...light, is another factor to consider. If your application requires longer periods of exposure, you’ll want...have shown that different fluorescent ligands can elicit varying degrees of β-arrestin-2 recruitment to ...descriptions, or technical manuals. Specific applications and matching dye/ligand choices For effective...beyond the diffraction limit of light. Dyes like silicon-rhodamine or Janelia Fluor (JF) dyes have specific...

...to disparate physiological effects and complicate any metabolic study. The Ralser lab has created a unique.... Seeliger MA, Young M, Henderso MN, Pellicena P, King DS, Falick AM, and Kuriyan J. Protein Sci. 2005...brief articles highlight the main features and applications of a partiular plasmid or set of plasmids. We...Wu Lab library can be found in the associated publication. Ma, et al. Cell Rep. 2015. PubMed PMID: 26190106...and ubiquitous reporter assay. In their recent publication, Kroeze et al. describe their enhanced TANGO .... Each gene was chosen to represent the primary splice variant in 38 types of human cancer. Many of the...Sanger sequencing, respectively. The speed and simplicity of GMAP allows scientists to easily design, generate...

... ITRs serve as the origin of replication and are required for replication, packaging, and vector persistence...viral rep (replication) and cap (capsid) proteins, which bind to the ITRs and initiate replication. ITRs are...binding site for Rep78 and Rep68 to initiate replication and a terminal resolution site that serves as... Rep proteins are important for viral genome replication, these mutations can affect transduction efficiency... of research is important for downstream AAV applications.  Tips & FAQs Due to their high GC content and... and hairpin structures, ITRs are prone to replication errors and disruptive mutations and truncations...mutations?  During propagation, mutants rapidly replicate in a growth culture and result in a high percentage...

...what we do not understand, and the complicated and unique metabolic processes of plants are being investigated...involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the biggest...difficulties that plant biotechnologists face is public perception of genetic modification (GM) and synthetic...benefits and future potential of the technology to the public in an open, responsible manner. At Revolution Bio...obstacles to plant synthetic biology In addition to public resistance, an overall limited understanding of...knockouts to better understand developmental, metabolic, and photosynthetic processes. Often the tools... intellectual property concerns – patents and licensing agreements. Developing a product without the use...

...constitutive splicing studies and has been optimized for high efficiency splicing. If intron splicing does not...potential applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves...in order to avoid complicating effects from global toxicity due to loss of splicing. To this end, destabilizing... system widely accessible for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol ...amenable to both high and low throughput screening applications. The system can be readily modified to use different...cloning platform for imaging and flow cytometry applications Designing customized fluorescent reporters can...luciferase reporter system for detection of pre-mRNA splicing   The Gideon Dreyfuss lab has developed rapid-response...

...host genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection... tools, thought was given on how to keep the replication of virus and its infectivity in check. Engineering...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety... be disposed of following your lab’s biohazard policies. Administering Virus Many virus experiments involve... accident-prone researchers. Note that safety policies will vary slightly from organization to organization...

...most transposable elements (TEs) results in the duplication of this target site sequence at the site of insertion...This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme...enzyme.  Class I TEs are considered replicative since every time they jump they make a copy of themselves...retroviruses both in their structure and mechanism of replication. They contain two genes: gag and pol. The pol...they move. Most Class II transposons have a non-replicative “cut and paste” mechanism of transposition: they... it integrates, which produces a target site duplication (TSD). Created in BioRender.com.   Autonomous...transposition activity  Tol2 transposon plasmids    Applications of transposons Now that you’ve learned about...

...many CRISPR applications, and they’ve also had success in clinical gene therapy applications. Read on to...to learn more about the current (and future) applications of lentiviral vectors! Lentiviral vector history...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...barcoding libraries have been used for complex applications like lineage tracing in heterogeneous tumors...developed integrase-deficient lentiviral vectors for applications that require the large genetic capacity of a...McGarrity GJ, Hoyah G, Winemiller A, Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl...

...industries. The cover letter, the first piece of an applicant’s writing read by any hiring manager or recruiter...ticket to a new job The first step in any job application is to pass the organization’s cover letter/resume...environment, the type, frequency and quality of publications, the techniques, and the lab's willingness to...papers, learning about the company or university policies, etc... Luckily, you can find most of this information... work on [phenomenon X], I  referred to your publications several times and I deeply admire your work....recruiter name], I would like you to consider my application for the [title of the position] at [name of the...achievements with measurable facts and do not forget to explicitly mention how you can contribute to the growth/...

...technique published every week! One of the newest applications is a colorful system that uses fluorescently...activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being transcriptional activation...characterizing the 4D nucleome. Future modifications and applications Ma et al. developed this technique using standard... et al. anticipate that the technique will be applicable to studies of cell cycle progression, epigenetics...to external stimuli. It could also have major applications to cancer, for example, the visualization of...Feb 23. Pubmed. Find the plasmids from this publication at Addgene. Dynamic imaging of genomic loci....2013.12.001. Pubmed. Find the plasmids from this publication at Addgene. A protein-tagging system for signal...

...Description Origin of Replication (ORI) DNA sequence which allows initiation of replication within a plasmid...plasmids are small circular pieces of DNA that replicate independently from the host's chromosomal DNA....virulence. All natural plasmids contain an origin of replication (which controls the host range and copy number...Minimally, lab-created plasmids have an origin of replication, selection marker, and cloning site. The ease... plasmids and the ability of plasmids to self-replicate within a cell make them attractive tools for the...plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing...broadly utilize plasmids across a wide range of applications. Some common plasmid types include cloning plasmids...

...Ligation Independent Cloning (SLIC) in a previous Plasmids 101 post. Although SLIC may be more cost effective...assembly meets CRISPR Gibson can be adapted to more complicated cloning schemes, such as those where the vector...CRISPR technology and is described in a recent publication by the Lockey lab (Wang, et al. 2015). In this..., Gibson assembly improves on two aspects of the SLIC methods. First, it uses a dedicated 5’ exonuclease...is added to repair the nicks in vitro, whereas in SLIC these constructs are repaired in vivo, which ends...ends up being much less efficient. In addition to SLIC and Gibson, there are yet more homology based assembly...CPEC (circular polymerase extension cloning) and SLiCE (Seamless Ligation Cloning Extract) to name two ...

...to be doing in humans; We have not had a broad, public conversation or consensus around the ethics of ... us with the second critique. Absent of a broad public consensus on genome editing, there was a generally...revisited at meetings when sufficient time, additional public fora for data review, and more lab research could...could be used to bring scientists and the public together to build consensus guidelines, but there was ...t have to look far for proof that this model of public involvement, research, and consensus can happen... separate non-governmental groups with seeking “public views on emerging IVF techniques designed to prevent...recognize so-called bad actors among them and “self-police” or whether the media and academic conference response...

... a bacterial origin of replication (ORI) will not allow for plasmid replication in mammalian cells, for... their bacterial counterparts, including how replication occurs and whether selection is necessary for...the nuclear genome, in the absence of plasmid replication, the foreign DNA will be degraded or diluted ...always mission critical to make sure the plasmid replicates in the cell, as many of these experiments are...expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr virus...contain an ORI since the integrated DNA will be replicated as part of the genome. Because the foreign DNA...Antibiotic Resistance Genes Plasmids 101: Origin of Replication All Plasmids 101 Articles ...

...range of applications. Image reproduced from Torres Cabán et al. 2022, under CC-BY-NC-ND license.   Find...Image adapted from Ino et al. 2022, under CC-BY license.   Find MTRIA sensors here: https://www.addgene.org...They generated variants optimized for different applications (v82 has higher affinity for glutamate but slower...reproduced from Liu et al. 2022, under CC-BY-NC license.   Find JEDI-2P plasmids here: https://www.addgene.org...ITAM to an RTK of interest and co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes...adapted from Farahani et al. 2022, under CC-BY license.   Find pYtags plasmids here: https://www.addgene.org...Image adapted from Koveal et al. 2022, under CC-BY license.   Find LiLac plasmids here: https://www.addgene.org...

...ITRs serve as a type of origin of replication to initiate replication of the dsDNA virus. Viral packaging...plasmid while the other plasmids with packaging or replication components are called helper plasmids. Below,... production, they can be helpful for certain applications and aid in ease of use. Our lentiviral guide...AAV are rep and cap. Rep is required for viral replication and also aids in infrequent genome integration...a helper plasmid and mediate efficient viral replication. More information on these viral genes is available...particles. E2A/E2B are responsible for viral replication, with E2B being the primary viral polymerase....is missing the endogenous E1 gene, making it replication deficient. This is typically supplied by the ...

...addition of a helper virus or plasmid allows it to replicate episomally, essentially remaining an extrachromosomal...extrachromosomal part of the host genome that can replicate and divide along with the endogenous DNA. AAV also...cell type doesn’t always lend itself to direct application of virus. However, the cargo capacity of AAV ...gamma-retro viruses. This is large enough for many applications, but in some cases multiple AAVs (co-infection...and AAV. If you’re wondering more about what applications these viruses are good for, we have plenty of...Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New...Gammaretroviral vectors: biology, technology and application. Viruses. 2011 Jun;3(6):677-713. DOI: 10.3390...