...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...

... 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...

...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...

...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...

...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...

...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...

... 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...

...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...