We narrowed to 320 results for: POR C
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TypeBlog PostPublishedSept. 14, 2021, 1:15 p.m...., it is important to not overgrow the plasmid at 37°C. Some researchers find that using a 30°C incubation...pnas.77.8.4847 Trepotec Z, Geiger J, Plank C, Aneja MK, Rudolph C (2019) Segmented poly(A) tails significantly...https://doi.org/10.2144/000112585 Ding T, Huang C, Liang Z, Ma X, Wang N, Huo Y-X (2020) Reversed paired-gRNA...Working on the scientific support team at Addgene, I get to see everything that can go wrong with a plasmid...
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Viral Vectors 101: Producing Your rAAV
TypeBlog PostPublishedJuly 16, 2024, 1:15 p.m....S., Croft, C. L., Futch, H. S., Ryu, D., Ceballos-Diaz, C., Liu, X., Paterno, G., Mejia, C., Deng, D.,...References Aurnhammer, C., Haase, M., Muether, N., Hausl, M., Rauschhuber, C., Huber, I., Nitschko, H...Esbin, M. N., Whitney, O., Walther, N., & Maurer, A. C. (2023). Transgene Expression in Cultured Cells Using...., Chtarto, A., Clark, K. R., Conlon, T., Darmon, C., Doria, M., Douar, A., Flotte, T. R., Francis, J....different titers are measured. A TCID50 measurement reports an infectious titer, meaning the concentration ... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog PostPublishedApril 5, 2018, 12:42 p.m....terminal); 2xNLS (C terminal) 47327 pET-28b-Cas9-His 6xHis (C terminal); NLS (C terminal) 62933...6xHis 6xHis (C terminal); NLS (C terminal) 62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS ...also made targeted insertions relatively simple in C. elegans and bacteria. But CRISPRing typical mouse...the stop codon! Figure 3: How to design a C-terminal insertion with Easi-CRISPR. Potential guide... homology arms and a selection marker, then electroporate it into mouse embryonic stem cells. After selecting...compatible with standard microinjection and electroporation methods using in mouse transgenics. Designing...between the ampulla and infundibulum. In vivo electroporation is performed immediately after injection. Image... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m.... standard strain, like DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity of ...freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting, the safest...ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus may appear as a...freeze-thaw cycles on hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with branched-DNA...that infect bacteria), many of which are doing important work in our microbiomes. Given that viruses are... and the type of virus, there have been varied reports of titer losses from 5% to 50% per freeze thaw ...vector: optimization of conditions for storage, transport and delivery." Japanese journal of cancer research... -
Hot Plasmids: Summer 2024
TypeBlog PostPublishedAug. 27, 2024, 1:15 p.m..... K., Chen, J., Pommier, G. C., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q.,... module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected with...Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of PRC2 with RvLEAMshort...glutaraldehyde crosslinking: representative micrograph (C), particle orientation distribution and Fourier-shell...tardigrades! Find RvLEAMshort here! Abe, K. M., & Lim, C. J. (2024). Small LEA proteins as an effective air-water...-02078-y. Anzalone, A. V., Gao, X. D., Podracky, C. J., Nelson, A. T., Koblan, L. W., Raguram, A., Levy...Serack, F., Harvey, J. W., Brauer, P. P., Pirtle, C. P., Coffey, A., Howard, M., Kamath, N., … & Weissman... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog PostPublishedJan. 24, 2023, 2:15 p.m....110, 073704 (2017). Hurt, R. C. et al. Genomically mined acoustic reporter genes for real-time in vivo ...visible in cells via TEM, and purified GVs under TEM. (c) Ultrasound imaging of tumor colonization by bARGSer-expressing...general method to create your own mARGAna cell line. (c) Z-projected phase contrast images of GV expressing...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used... to study gene expression in cells. But these reporters have limited use in vivo because they are based... penetration depth as well as high spatial and temporal resolution, making it ideal for visualizing gene...expression in vivo. But what ultrasound lacked was reporter genes that could produce ultrasound contrast. ... -
Developing a cold tolerant E. coli using specialized chaperones
TypeBlog PostPublishedJan. 28, 2020, 2:35 p.m....defined between a fairly nippy 20°C to a decidedly uncomfortable 45°C). Their more adventurous cousins ...of growth, say around 37°C, is a far cry from their native conditions of ~0°C, which drastically affects... But what about placing the mesophilic hosts at 0°C? That won’t work out either - most mesophilic systems...severely cold conditions. To understand their unique importance, let’s begin with a few basics. Mesophiles vs...understanding and appreciating their biology. The importance of psychrophilic enzymes From an industrial perspective...decided to create a hybrid system where we could incorporate cold-tolerance inducing genes within a regular...scientist from Addgene. The talk highlighted the importance of reagent repositories and the myriad facilities... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....into the A2 noradrenergic area of the caudal NST. (A-C) Low-magnification images showing viral spread. (D-F...Backer, M. W. A., Brans, M. A. D., Luijendijk, M. C., Garner, K. M., & Adan, R. A. H. (2010). Optimization...Gruntman, A. M., Su, L., Su, Q., Gao, G., Mueller, C., & Flotte, T. R. (2015). Stability and compatibility...data free. Nature, 5–7. O’Connor, D. M., Lutomski, C., Jarrold, M. F., Boulis, N. M., & Donsante, A. (2019...Malaker, S. A., Poweleit, N., Maynard, L. H., Adams, C. M., Leib, R. D., Cirolia, G., Thomas, D., Stamnes...doi.org/10.1016/j.ymthe.2005.02.021 Zuk, P., Sanchez, C. E., Kostick, K., Torgerson, L., Muñoz, K. A., Hsu...experiments didn’t provide any information on other important factors such as what is the best promoter to use... -
Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools
TypeBlog PostPublishedDec. 6, 2018, 1:34 p.m....construct for C. elegans gene knockdown Article contributed by Alyssa Cecchetelli Listen to the C. elegans...segment RNA interference (RNAi) is extensively used in C. elegans research to study gene function. RNAi is ...commonly achieved by feeding E. coli expressing dsRNA to C. elegans. These dsRNAs target specific mRNA transcripts...previously not an option, including Drosophila and C. elegans. Find the TurboID plasmids here! Branon...producing long unspecific RNA fragments that are incorporated into the dsRNA necessary for RNAi. The Vellai...adjacent to the T7 promoters to prevent the incorporation of non-specific nucleotides into the dsRNA leading...enzyme and a small-molecule substrate to map the spatiotemporal distribution and interaction networks of specific... -
Degrading DNA with Cascade-Cas3
TypeBlog PostPublishedFeb. 11, 2025, 2:15 p.m...., Banh, L. M., Williams, C., Toh, M. S., Irby, M. J., Edwards, L. S., Lin, C., Owen, A. L. G., Künne, ...contains all five of these Cas proteins, while type I-C Cascades do not contain Cas6. Regardless of the subtype...Resources References Loeff, L., Brouns, S. J., & Joo, C. (2018). Repetitive DNA reeling by the Cascade-Cas3...Structural snapshots of R-loop formation by a type I-C CRISPR Cascade. Molecular Cell, 83(5), 746-758.e5....Coons, M. M., Klompe, S. E., Lied, A. M., Smith, S. C., Vidal, B., Donohoue, P. D., Rotstein, T., Kohrs,....com. The Cascade complex Cascade is an important component of the CRISPR Cas3 system. Most Cascade... appears to be conserved. This DNA degradation portion can be bypassed by leaving out Cas3 — a tactic ...