...scatter (FSC) and provides information about the relative size of the cell; larger cells produce more FSC...experiment, you would need a way to determine the relative contribution of yellow and orange from FITC and...interest with a fluorescent reporter on the N- or C-terminus of the protein. One downside of fusion proteins...doi.org/10.1002/cpim.40 Picot J, Guerin CL, Le Van Kim C, Boulanger CM (2012) Flow cytometry: retrospective...intensity of events. Since intensity positively correlates with expression, the further the shift of a cell...

...high Cas9 MOI considering that A549 cells are relatively easy to transduce. I ended up with too many Cas9...on genomic DNA. The surveyor assay itself was relatively easy. Figure 4: Detection of edited BRAF... new scientific observations. References 1. Rago C, Vogelstein B, Bunz F. Genetic knockouts and knockins...Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Nat ...

... P., Rodriguez-Aguayo, C., Walton, B. L., Sood, A. K., Greaver, J., Janssen, C. F., & Lopez-Berestein,... Academic Press. Turner, P. V., Brabb, T., Pekow, C., & Vasbinder, M. A. (2011). Administration of substances...ingestion, such as experimental therapies or pain relief, straight to the stomach. This technique involves...

...assembly of NgAgo/ssDNA requires incubation at 55 °C - a dangerous, non-physiological temperature for mammalian...genome or just after a stop codon (as required for C-terminus epitope tagging). NgAgo survey Since it was..., he tries random things (101 lessons on Spanish, C++, guitar, photography) and often dreams of a sunny...expression plasmids. Cas9 guide RNA delivery does however rely on cloning into a vector or in vitro synthesis, ...

...for Tet transregulators. Krueger C, Pfleiderer K, Hillen W, Berens C. Biotechniques . 2004 Oct;37(4):546...developed. tTA was created by fusing tetR with the C-terminal domain of VP16 (virion protein 16), an essential...encoded tetracycline resistance. Hillen W and Berens C. Annu Rev Microbiol . 1994;48:345-69. PubMed . Transcriptional...cells. Yao F, Svensjö T, Winkler T, Lu M, Eriksson C, Eriksson E. Hum Gene Ther . 1998 Sep 1;9(13):1939...biomedical research. Bockamp E, Maringer M, Spangenberg C, Fees S, Fraser S, Eshkind L, Oesch F, Zabel B. Physiol...rTetR, which reversed the phenotype and created a reliance on the presence of tetracycline for induction,...

...References and Resources References Radtke, A. J., Chu, C. J., Yaniv, Z., Yao, L., Marr, J., Beuschel, R. T.... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv...the IBEX Knowledge-Base is named on the Zenodo release in keeping with their value of shared ownership...

....2019.02.037 Kagiampaki, Z., Rohner, V., Kiss, C., Curreli, S., Dieter, A., Wilhelm, M., Harada, M., Duss...References and resources References Feng, J., Zhang, C., Lischinsky, J. E., Jing, M., Zhou, J., Wang, H.,...inhibitory feedback in response to norepinephrine release, and clinically-used drugs targeting these receptors...

...site for a total of 2 binding sites per antibody. C) IgG’s are big enough that multiple labels (green ... IgG secondary antibody (Pleiner et al figures 4A-C). And since they have limited cross-reactivity between...Bates, M., Böhning, M.I., Chug, H., Görlich, D., Lee, C., Pleiner, T., Schliep, J.E., Stark, H., Trakhanov... can be cleaved by the bdNEDP1 protease, thus releasing purified nanobodies from the column. bdNEDP1 protease...

... Quiñones-Valles, C., Espíndola-Serna, L., Martínez-Antonio, A., Quiñones-Valles, C., Espíndola-Serna,... A., Gupta, R., Dyson, K., Rao, P., & Leonard, A. C. (2018). Origin recognition is the predominant role...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...replication independently are said to be under relaxed control, as they only require the host's replication...replication (oriC), where initiation begins. This process relies on the initiation protein DnaA. First, DnaA must... chromosomal DNA ensure that the helicases are released from the DNA once the replication forks reach ...

...In summary, this cell free system allows for a relatively simple, controlled “one-pot” reaction capable... system is capable of robust expression of high G+C content genes, as validated through monitoring the...

...10.1126/science.1206593 Powers S, Miranda C, Dennys-Rivers C, Huffenberger A, Braun L, Rinaldi F, Wein....2020.107878 Sinnett SE, Hector RD, Gadalla KKE, Heindel C, Chen D, Zaric V, Bailey MES, Cobb SR, Gray SJ (2017... expand as Rett scientists deposit their MECP2-related plasmid tools with Addgene and share additional...Methyl-CpG-binding protein 2 mutations and phenotype-genotype correlations. Am J Med Genet 97:147–152 . https://doi.org...Determines the Clinical Course of Rett Syndrome and Related Disorders. Cell 152:984–996 . https://doi.org/10.1016...

...System, AmpR 26100 pFHMSP-LIC-C N/A Expression vector for secreted proteins with C-terminal hexahistidine tag...dimensional structures of proteins of medical relevance, and place them in the public domain without restriction...TEV cleavage, Bac-to-Bac Expression System, AmpR Related Resources SGC: http://www.thesgc.org/ SGC Toronto...

...Number (A) Year Published (B) Number of Citations (C) Journal Name (D) Percent of Papers without Addgene...Journal (F) Weight (=F/E) (G) Weighted Citations (=C*G) 1 2010 3 Nature  3.3% 10.5%   3.2  9.6 ...demand for the related plasmid. Under such circumstances, we would observe this same correlation, but it wouldn...pulled the journals from 2005 to 2015 in areas most relevant to plasmids: Genetics & Heredity, Cell Biology...aggregated this much, you can see the 2013 ‘bump’ related to the CRISPR/Cas papers and Addgene deposits, ...Addgene, in contrast to other reasons for the correlation. However, determining this exactly is difficult... remaining difference caused by depositing the related plasmids with Addgene? Probably not all of it. ...

...Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) ...Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung... https://doi.org/10.1126/science.aaq0179 Myhrvold C, Freije CA, Gootenberg JS, Abudayyeh OO, Metsky HC...demonstrated that SHERLOCK could reliably distinguish between Zika and a closely-related virus, Dengue from multiple...simple, portable, and inexpensive platforms to reliably detect nucleic acids at the atomolar level. The... easy to transport and work rapidly, providing reliable results in as little as an hour. Find the SHERLOCK...from cell-free DNA fragments as well as health-related single nucleotide polymorphisms (SNPs) from human...

...be performed on all relevant bases within the editing window, for example all C’s will edit to T’s. Thus... single base pair edit. (Cytosine deaminases edit C-to-T and adenosine deaminases edit A-to-G.) In short...takes place on the free strand of DNA and converts a C to U without producing a double strand break. ...

...mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments in mammalian...can form it’s chromophore at a more comfortable 37°C (5). Dendra2 photoconverts from green to red fluorescence... pH stability, chromophore maturation rate at 37 °C, turnover rate, and the level of background fluorescence...: 1035-1040. PubMed PMID:18724362. 8.Stiel, Andre C., et al. "Generation of monomeric reversibly switchable...resolution imaging, where the power of the technique relies heavily on the same fluorophore being imaged multiple...photons achievable with photoswitchable dyes. Correlative light and electron microscopy (CLEM) is an imaging...

... a given disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged... Since the bacteria are subsequently grown at 37 °C, the recombineering plasmid is not maintained. Recombination...directed mutagenesis methods Current SDM methods rely on in vitro assembly. To make a point mutation, ...some of which are proprietary. These methods also rely on ssDNA ends to match up regions of homology, so...better one. Trehan et al. wondered if, instead of relying on in vitro assembly, they could make the E. coli...

...may have an entirely different use in mind! This section of our plasmid pages is relatively new, so some...here. Articles Citing this Plasmid: This is a relatively new feature of our plasmid pages, and provides...listed according to whether they’ll end up on the N or C terminus of the transcribed and translated insert....

...common fluorescent proteins and their mutations relative to wildtype GFP. Although not listed here, many... fluorophores Fluorescent Protein Mutations Relative to Wildtype GFP EGFP F64L; S65T EYFP S65G;...the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist ...reported in 1996, is an eleven β-sheet-containing “barrel” shape, with the chromophore concealed at the center...spectrum are not GFP derivatives, but are instead related to the dsRed protein isolated from Discosoma sp...

...constructs 69570 and 69571 together encode the N and C terminal segments respectively of a GFP dependent ...-N-CretrcintG Connie Cepko 69571 AAV1 pAAV-EF1a-C-CreintG Connie Cepko 75469 AAV1 pAAV-EF1a-Flp-DOG-NW...are some of the AAV and lentiviral preps we have released in the last few months: Lentiviral preps for ...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality...