...., Pan, C. Y., Rizzo, K., Bertumen, J. B., Morales, C., Oluniyi, P. E., Nguyen, J., … Chiu, C. Y. (2023...36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval, A., Harris-McCoy, K. C., Hensley, S. E., Zhou, D., Lin..., Iparraguirre, A., Lopez-Camacho, C., Wherry, E. J., & Ertl, H. C. J. (2007). Adenoviral vectors persist...Borkenhagen, L. K., Fieldhouse, J. K., Seto, D., & Gray, G. C. (2019). Are adenoviruses zoonotic? A systematic review....org/10.1016/B978-0-12-801238-3.02523-X. Hofmann, C., Löser, P., Cichon, G., Arnold, W., Both, G. W., ...AdVs are ubiquitous (Schmitz et al., 1983), yet rarely cause severe disease in humans (and no vaccines...stages of infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(...

...Shaking incubator at 37 °C Stationary incubator at 37 °C Water bath at 42 °C Ice bucket filled with ice... storage at 4°C and let warm up to room temperature and then (optional) incubate in 37°C incubator. Mix...also make their own competent cells. This is a relatively simple procedure and is useful for performing...transform Procedure Take competent cells out of -80°C and thaw on ice (approximately 20-30 mins). Remove...placing the bottom 1/2 to 2/3 of the tube into a 42°C water bath for 30-60 secs (45 secs is usually ideal...without antibiotic) to the bacteria and grow in 37°C shaking incubator for 45 min. Pro-Tip This outgrowth...and won't grow in colonies. Incubate plates at 37°C overnight. Tips and FAQ How can I save time when carrying...

...Optimized mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene disruption in M. abscessus...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs.... E., Young, D. F., Goswami, K. K., & Russell, W. C. (1987). Isolation and characterization of monoclonal...39406745. Optimizing mScarlet tags for live imaging in C. elegans By Alyssa Neuhaus Red fluorescent proteins...maturation rate limits its use in systems like the C. elegans embryo, where developmental processes happen...recently generated new constructs specifically for C. elegans (Cao et al., 2024). The team designed coding...mScarlet3 and mScarlet-I3 that were codon-optimized for C. elegans and included artificial introns to improve...

...jcm.23.5.959-961.1986 Lobstein, J., Emrich, C. A., Jeans, C., Faulkner, M., Riggs, P., & Berkmen, M. (2012... some people still use amp? Carb is expensive (relative to amp), and for some applications, the benefit...References Hwang, J. M., Piccinini, T. E., Lammel, C. J., Hadley, W. K., & Brooks, G. F. (1986). Effect...their mode of action and have a very similar relationship as amp and carb (they are interchangeable). ...low rate, there is evidence that zeocin is never entirely detoxified, even when Sh ble is expressed (Trastoy...

...T-regs) into their microenvironment by producing C-C motif chemokine 22 (CCL22) and TGFβ. T-regs are suppressive...tolerance-producing antigens enunciated above. Finally, signals released from the microenvironment help create tolerance...eradicate metastasis or eliminate the probability of a relapse. Chemical and radiation therapies cause a transient...

...CyPet with C-terminal His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus...YPet with C-terminal His tag SYFP2 Yellow Mammalian Express a gene of interest fused to the C-terminus...Mammalian Express a gene of interest fused to the C-terminus of monomeric Cerulean mTurquoise2 Cyan ...Mammalian Express a gene of interest fused to the C-terminus of Amber mVenus N1 Yellow Mammalian Express...Mammalian Express a gene of interest fused to the C-terminus of monomeric Venus pCEP4YPet-MAMM Yellow...Mammalian Express a gene of interest fused to the C-terminus of LSSmOrange mRuby2 Red Mammalian Expresses...FRET occurs. By photobleaching the acceptor, you release the donor's quenching and the fluorescence of the...

...parameters: Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Enzyme Activation 95 10...titer: $$T = {V*C*D\over v}$$ Where: T = Infectious titer, TU/mL V = Viruses per genome C = # Cells per ... across the dilutions. RPP30 copies should be relatively constant across samples. In the RRE example below...concentration of RPP30 positive droplets stays relatively even across samples (green). To increase the ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable...Benzonase in DMEM complete to each well. Incubate at 37 °C for 30 min. Gently aspirate media from wells, using...can be used for ddPCR immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples...

... D. W., Kumaran, R., Rego, M., Schofield, C., … Laflamme, C. (2024). A consensus platform for antibody... H., Laflamme, C., Houston, D. W., Polinski, N. K., Meijers, R., Levey, A. I., Saper, C. B., Errington...Ayoubi, R., Ryan, J., Gonzalez Bolivar, S., Alende, C., Ruiz Moleon, V., Fotouhi, M., Alqazzaz, M., Southern... strides in making science more accessible and reliable. We continue to expand our educational content...

...Bryson, C., Fogg, M., Baker, M. P., Herskovitz, J., Goletz, T. J., Zhou, L., Moxness, M., Flynn, G. C., Narhi...trained on vast amounts of data to calculate the relative immunogenicity score (Mattei et.al., 2022).    ...Antidrug Antibody Formation in Oncology: Clinical Relevance and Challenges. The oncologist, 21(10), 1260–1268...

...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....serotype, as some rAAV serotypes are abundantly released into the culture media, while others remain cell-associated...Wilson, J. M. (2010). Efficient serotype-dependent release of functional vector into the culture medium during...

.... K., Chen, J., Pommier, G. C., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q.,...representative micrograph (C), particle orientation distribution and Fourier-shell correlation curve (D), and the... 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...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...

...changes from C•G-to-T•A are mediated by cytosine base editors (CBEs) and base changes from A•T-to-G•C are mediated...called point mutations that change C to T, G to A, A to G, and T to C, represent about two-thirds of known...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464...Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR (2020...PACE and PANCE systems use M13 bacteriophage, which rely on the expression of gene III to create infectious...

...comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant, DsRed.T3, that’s ...quenched in acidic environments, while mRFP is relatively stable and fluorescences even at the acidic pH... GFP/RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence suggests an increase in autophagic...autophagic flux, while an increase in GFP fluorescence relative to RFP fluorescence suggests a decrease in autophagic...∆G, however, can not be conjugated to PE due to a C-terminal glycine deletion, so it remains in the cytosol...changes in autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this...important to remember that because Keima’s fluorescent relies on lysosomal acidity, this biosensor can’t be used...

...transgenic nematodes, including C. elegans and C. briggsae. Other related miniMos plasmids can be found ... below lists plasmids that contain a worm ( C. elegans, C. briggsae , etc.) gene or sequence: ID Plasmid...Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses...General Tools Fire Lab C. elegans Vector Kit - Andrew Fire Lab. A set of vectors for C. elegans research, ...distributes stocks of C. elegans. Silencing Genomes - Cold Spring Harbor's guide to RNAi in C. elegans WormAtlas...concerning the genetics, genomics and biology of C. elegans and related nematodes. WormBook - A comprehensive, ...peer-reviewed chapters covering topics related to the biology of C. elegans and other nematodes. Wormbuilder...

...specific genomic locus: C-BERST Article contributed by Klaus Wanisch  Listen to the C-BERST podcast segment...processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA. 2018 PubMed PMID:29109157. Nanobody ...Sontheimer lab used this technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements...in future studies of other genomic loci. Find the C-BERST plasmids here. Gao at al. Nature Methods (...protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find all of the nanobody toolkit ...

...construct [1,2], i.e. positioning of the FP sequence relative to cell compartment targeting sequences, which...to characterize FP-fusion proteins functionally relative to the untagged protein of interest. To avoid ... retrieval sequence only functions at the extreme C-terminus [3]. You must therefore determine your cloning...strategy and decide whether you will create an N- or C-terminal fusion based on your experimental needs. ...

...response gene c-fos. For example, using Chr2 stimulation should result in robust increases in c-fos expression...stimulation compared to controls without Chr2. c-Fos is a reliable and widely used molecular marker of neuronal...proceed with your behavioral experiments. Comparison of c-fos amongst animals that receive different types of...neuronal activation, but it may still be possible to use c-fos expression to get the evidence you need. For example...activation in the novel context as determined by reduced c-fos expression, thus showing successful inhibition... as non-human primates [7, 8], zebrafish [9], and c. elegans [10]. If used properly, optogenetics is sure...literature to find all the applications of optogenetics relevant to your field. Ultimately, your question should...

...showed that iGluSnFR could detect neuronal activity in C. elegans, zebrafish, and mice. Second Generation ...Improved trafficking and localization through different C-terminal anchoring domains The original iGluSnFR in...adapted from Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane domain ...mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol anchor. SGZ contains...domain, followed by a modified form of the cytosolic C-terminal domain of Stargazin including a terminal ...give researchers the ability to monitor glutamate release from neurons and other brain cells in vivo. Glutamate...differentiating between synaptic vs. extrasynaptic glutamate release. Building on the previous generation of the SF-...

... 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...restriction enzyme digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to ...reducing the culture volume into which the virus is released, a more concentrated virus solution can be obtained...

... present, it will repress expression of mKate2 relative to EBFP2. Weiss’s group used these reporters to...chain of BoNT/B into two fragments (BotNT-N and BotNT-C) and joined each fragment with one part of an established..., and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids at Addgene... it inactive, thus inhibiting neurotransmitter release. How is this light-mediated? The researchers split...visualizing the location and function of microbes rely heavily on fluorescent protein genes. These optical...