... Their focus at the lab is on plant-pathogen interactions, and her aim is to produce constructs and edit...biosensors designed to elucidate the molecular interactions that go on between plants and their pathogens... share and educate and enable plant science in general. We are funded by a charity and since we are making...elements anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...piece of DNA. We do a lot of protein-protein interaction work and we’ve gotten some tools back from Addgene...Twitter account recently inspired by an academic in literature who gave a talk to the women’s research network...

... both vector amplification with low fidelity polymerases and the cost of synthesizing longer oligos, which...using the same three step protocol:  Fragment generation - Any linear DNA fragments can be assembled in... the template binding region, with a melting temperature (Tm) of around 60 °C for all primers. The length...simple cloning procedures. Use high fidelity DNA polymerase This will minimize random errors during PCR: ...aliquots of PCR master-mix, containing buffer, polymerase and dNTPs, can be stored frozen: just defrost...mutagenesis by placing homologous ends on DNA using polymerase chain reaction." Biotechniques 10.1 (1991): 62...

... Major restaurant chains including Chipotle and Panera Bread have pledged to eliminate the use of antibiotics...number of phagemids would be needed for an in vivo therapy. Another strategy for CRISPR antimicrobials combines...replication-competent phages and antibiotics. Phage therapy fell out of fashion with the introduction of antibiotics...Rodolphe Barrangou. “A CRISPR design for next-generation antimicrobials.” Genome Biol. 15(11):516. (2014...Ido, Miriam Manor, Ruth Kiro, and Udi Qimron. “Temperate and lytic bacteriophages programmed to sensitize... phage therapy...

...reporter enzyme. For multicolor staining, careful consideration must be used when selecting which antibodies...meticulously screening, optimizing, and characterizing several anti-mouse and -rabbit nanobodies. If you’re interested...described in Pleiner et al, the key steps for generating nanobodies are outlined below; no alpaca required...due to their N-terminal His14-bdNEDD8 tag. After several washes, this tag can be cleaved by the bdNEDP1 ...nanobodies from the column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: ...Maleimide-conjugated HRP can be found here and several maleimide-conjugated dyes are also commercially...

...infection; however this would not be great for a gene therapy vector! For viral vectors, only the protein for...viral vector particles are made while missing the polymerase gene may seem like a mystery. But it’s simple...transcription and translation. Here, we will discuss several aspects of the viral “life cycle” in order to understand...step 3), but note also that it is worthwhile to generalize the idea of “transduction” to mean any transfer...et al., 1973). Interestingly, retroviruses are generally only “pseudodiploid” in that — while they have... M., & Bishop, J. M. (1973). RNA-directed DNA polymerase of Rous sarcoma virus: Initiation of synthesis...

...incorporated a discovery-based lab on planarian regeneration into one of my own courses, I was excited to...phosphorylation on the enzyme’s structure, function, and interaction with other proteins.   It is important to...be authentic – students should be attempting to generate new knowledge that will be of genuine interest...and bench work to identify regions of MDH that interact with citrate synthase and presented their findings...increasing diversity and inclusion in STEM disciplines (Bangera & Brownell, 2014; Duboue et al., 2022), as they...classroom.   References and Resources References Bangera, G., Brownell, S. 2014. Course-based undergraduate...

...deliver all the protein and RNA components, and several sgRNAs must be screened to identify effective ones...Schematic of the Cre-dependent SPH construct for generating the SPH mouse. (B) AAV plasmids used to express...gene expression. Gene isoforms refer to mRNAs generated from the same gene but with different untranslated...regulation — at least, more straightforward than generating cDNA for each gene isoform individually. To highlight...their genomic DNA sequenced. Guide RNAs targeting several known and some unknown regulators of neuronal differentiation...Although cDNA overexpression approaches may still be preferable for individual gene activation in select in vitro...

...ricin toxicity genes. The screen also discovered several genes previously not associated with ricin regulation...known ricin regulation genes compared to results generated using non-targeting guides as controls. While ...sites. Results from the screen also reproduced several previously observed characteristics that influence...activity: 1) mismatches closer to the PAM are less tolerated than mismatches more distal to the PAM, and 2)...off-target activity, Morgens et al drew some generalizable conclusions about off-target activity, demonstrate...Michael C., et al. "A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility...

...genotypic and phenotypic artefacts. The life science literature is a litany of false interpretation in cell culture...Down-Regulation of HIF-1α by NDRG2 Expression Attenuates Proliferation and Invasion in Renal Cancer Cells, Marker-independent...out by the commercial repository. Repositories generally ensure cell line identity and purity making it...to the original as possible and so any results generated are reproducible and reliable. When you order ...which can be stored in liquid nitrogen long term, preferably some of these vials will be stored in a separate...identity. The Nature Publishing Group recently reiterated its strict stance on the matter. Funding bodies...

... to write, to read efficiently, to search the literature, and to synthesize a large volume of information...appreciate the discussion sections of the primary literature as a resource that both summarized results and... sure if I should cite reviews or the primary literature or both. I ended up citing both because I used...the scope and the content. Submit your article! Overall, writing a review can be overwhelming and challenging...Thompson, and Thomas D. Gilmore. "Histone acetyltransferases and histone deacetylases in B-and T-cell ...Addgene Blog Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing Tips for Improving...

...circuits and dynamics. FPs are generally bright, stable and well tolerated by cells. They can be used for...neither disturb POI folding nor its ability to interact with protein partners within the cell. However...can affect protein folding and protein-protein interactions. Also, overexpression of FPs can trigger protein...ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with facile ...monster” fluorescent proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags...

...was first developed in 2001, with successive generations developed ever since. GCaMP6 was offered through... added jGCaMP7 and eventually jGCaMP8, later generations of GCaMP6, to the service. While they were both...jGCaMP8 per year from 2016–2023. All three generations are regularly requested and a valuable part of...size of the incremental improvements between generations: jGCaMP8 allows for the “picture” of a train ... former approach and involves reading primary literature, soliciting information from requestors and depositors...Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

...soon find yourself encountering flow data in the literature. Data reported from flow cytometry experiments...work is done in flow cytometers, and output is generated through measurement of events. An event is something... machine is plotted as a dot on an X,Y axis to generate a dot plot (Figure 1). Clustered events are called...                         Figure 1: While the plot generated often looks like the graph on the left, it might...intersection of X,Y. For flow plots, however, that is generally not true. Because of the variations in fluorescent...Of course, this is quite easy to do on a plot generated in Biorender! While a helpful visualization tool...

...Bert Vogelstein has published their method for generation of recombinant adenoviruses: A Practical Guide...

... through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start The health of the HEK293T ...

...member at Albert Einstein College of Medicine. Several sets of near-infrared fluorescent proteins (NIR...the NIR light coming from your source and to your camera.  Two-photon image of miRFP670nano3-expressing...reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of a probe leads to less interference...al., 2018). For detection of protein-protein interactions, we recommend using NIR split reporters based...detect spatiotemporal dynamics of protein-protein interactions. Nat Methods 11, 641-644 (2014). Filonov, G....reporter for in vivo imaging of protein-protein interactions. Cell Chem Biol 20, 1078-1086 (2013). Shcherbakova...

...candidate regulatory kinase is known, it’s possible to generate non-phosphorylated recombinant protein and perform...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...position. Practical tips for your experiments To generate as much pSer-incorporated protein of interest ...be optimized for parameters such as induction temperature and length of induction, which will vary between...between different proteins. Beyond these standard considerations, it is important to determine how much phosphorserine... effects of pSer on enzyme function, protein interaction, etc. by comparison to non-phosphorylated or ...

...that only 11% of the academic research in the literature was reproducible by their groups. Finally results...copes with the growing amounts of digital data generated in labs. While many labs approach the situation...are available on the market today, ranging from general note taking tools, to more scientific field-based...From pictures to excel sheets, researchers today generate vast amounts of digital data during their work...within your lab? In every lab, there are certain hierarchies and restrictions to data access. If these are...walk you through your ELN’s features and this interaction will give you an idea of how responsive and supportive...

...suggests there are hundreds. This only includes general lab strains designed for subcloning or protein ...ATPase that is essential for recombination and general DNA repair Reduces plasmid recombination, increases...Δleu mtl1 DH10B Streptomycin MC1061 derivative. General cloning and storage, blue/white screening, leucine...)7697 mcrA Δ(mrr-hsdRMS-mcrBC) λ- DH5alpha   General cloning and storage of common plasmids, blue/white...galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning and plasmid maintenance, blue/white screening...(TetR) TOP10 Streptomycin MC1061 derivative. General cloning and storage, blue/white screening. F- mcrA...

...produce molecular biological tools like Taq polymerase, T7 polymerase, EcoRI, and various fluorescent proteins... elucidating the molecular mechanisms behind interactions between crops like potatoes and their microbial...that enable E. coli and Citrobacter freundii to generate sugars from cellulosic biomass. Dr. Duedu would...construction of new plasmids. Dr. Duedu will also use polymerases produced using plasmids from Addgene to test ...