We narrowed to 419 results for: SAM-2

...//doi.org/10.1002/1096-8628(200022)97:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, ...Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive...X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https://doi.org/10.1038/13810... (2000) Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype correlations. Am ...https://doi.org/10.3389/fgene.2019.00625 McGraw CM, Samaco RC, Zoghbi HY (2011) Adult Neural Function Requires...

...   Figure 2: Synthetic promoters can be paired for targeted expression. This same pairing of synthetic...retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved...1345-1356. https://doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression...different synthetic promoter driving expression of the same transgene: an optogenetic reporter, channelrhodopsin...in mice is not a good predictor for targeting the same cell class in NHPs (R=0.38) and humans (R=0.32),...), but the odds of that promoter targeting the same cell class in all three species are still greater ...in monkeys was a good predictor for targeting the same cell class in humans (R=0.67).  A database of the...

...phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the importance of... the open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...optimized Sep-OTSλ and can recommend its use (6).  2. C321, a specialized bacterial host strain for pSer-incorporation...biology4.5 (2002): E127-E130. PubMed PMID: 11988757. 2. Hornbeck, Peter V., et al. "PhosphoSitePlus, 2014... purified from the system, researchers can run samples on SDS-PAGE gels containing Phos-Tag, a small molecule...non-phosphorylated serine at the TAG codon of the same expression vector used for phosphoserine incorporation...incorporation of serine) population of protein using the same expression plasmid. This is a particularly important...

...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...briefly boiling the sample before use or by directly adding a small volume of the sample to the PCR reaction...enzyme digest; however colony PCR can accomplish the same thing in less time and for less money. The key steps...also great for screening clones created with the same backbone but that contain different inserts. When...sequence of the insert is, allowing you to use the same primer pair to screen for the presence of many different...

...transcription factors 1 and 2. See Aranda-Díaz et al. for more information. The El-Samad lab predicts that this...difficult-to-manipulate primary cells, achieving knockdown within 1-2 hr of reagent delivery. TRIM-Away is suitable for ...factor 1 controls expression of transcription factor 2, which directly controls expression of a reporter ...Factors Podcast Segment Researchers from the El-Samad lab recently deposited plasmids encoding synthetic...encoding these synthetic transcription factors, the El-Samad lab has deposited plasmids encoding fluorescent ...

... Multiple Introns.” Plant Communications, vol. 2, no. 2, Elsevier BV, Mar. 2021, p. 100135. Crossref, ...Constructs.” PLoS ONE, edited by Jean Peccoud, vol. 6, no. 2, Public Library of Science (PLoS), 18 Feb. 2011, p... vol. 5, no. 10, American Chemical Society (ACS), 2 May 2016, pp. 1059–69. Crossref, https://doi.org/10.1021...of Plant Genomes.” The Plant Genome, vol. 16, no. 2, Wiley, 9 Mar. 2023. Crossref, https://doi.org/10.1002...Crossref, https://doi.org/10.1186/s12870-020-02388-2. Stuttmann, Johannes, et al. “Highly Efficient Multiplex...Crossref, https://doi.org/10.1038/s41598-019-46171-2. Resources on Addgene.org Modular Cloning (MoClo...unique genomic targets or multiple sgRNAs against the same target to ensure efficient cutting. Beyond sgRNAs...

...purify IVT sgRNA in the lab. Figure 2: Cas9 RNP complexed with 2-part cr:tracrRNA tags genes more effectively...into stem cells. Cells are allowed to recover for 2-5 days and then analyzed for tagging efficiency using...IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient...cell suspension, transfect and let cells recover for 2-5 days.Note: We use the Nucleofection method for delivery...success was ~%50. If you get low knock-in efficiency (<2% positive by ICC for the tag) or do not see any cells...PMID: 28096221. PubMed Central PMCID: PMC5312033. 2. Dewari PS, Southgate B, Mccarten K, Monogarov G, ...turnaround time 5-7 days); because the tracrRNA is the same in all csRNP experiments, it can be ordered in bulk...

...DNA and processed the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly analyzed ...look for potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual... mean the sample is contaminated? No - what really matters is the number of hits to the same sequence ...these known impurities, as was the case for clean sample AAV-44362. In addition to these expected hits, ...sequence (gene). In a clean sample, there are typically fewer than 10 hits to a given sequence, and we believe... the contaminant we mixed into the original AAV sample at 5%. Detecting 5% contamination is a good start...low we could go. We prepared a range of spiked samples from 0.1-20% and discovered that, with a high enough...

...are many docking sites available for integration. 2. How soon would you like your flies ready for use?...beautiful immunofluorescent images like in this paper (2). Metabolic assays such as Seahorse can be employed...PMID: 16547094. PubMed Central PMCID: PMC1526508. 2. H. Jiang,  et al. Cytokine/Jak/Stat signaling mediates...recombination so that the transgenes can be inherited on the same chromosome. Targeted Insertion allows for direct...variants of YFG because the transgenes are found in the same locus across fly lines (i.e. there won’t be positioning...

...required to make a plasmid. In 2023 (and at closer to 2 million plasmids shared than not!), we think it’s ...time does it take to make a plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n...go wrong, replace them for free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether...overall inflation of USD has gone up 19.8% in the same time period. For ease of calculations*, let’s split...inflation at… roughly 34.9%, which means that the same plasmid would cost $155.25 in reagents and consumables...

...by a factor of 2. This means that if you set up two identical assays with the same sample, you could end...amplitude measured for each droplet in each well (Figure 2). A clean ddPCR should have a clear separation between...as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has ...standard curve is giving an accurate readout of your samples.  Of course, if qPCR were perfect there would be...must serially dilute your virus before adding your sample to a mastermix. At Addgene, we typically load 3... a wide range of titers. We usually dilute our samples 1:6 million to 1:25 million.  After making the ...and droplet oil that you use should come from the same manufacturer. Otherwise you will end up with poor...

...or non-homologous end joining (see Figure 2).  Figure 2: Schematic representation of integration...due to insertional mutagenesis (Howe et al., 2008). 2. Transient expression of transgenes/shRNAs If you ...vectors: progress and applications. Gene Therapy, 17(2), 150–157. https://doi.org/10.1038/gt.2009.135. PubMed...efficiently during infection. Journal of Virology, 70(2), 721–728. https://doi.org/10.1128/JVI.70.2.721-728.1996...performing quantitative PCR titration of vector DNA in samples harvested 24 hours post-transduction (Wanisch and...

...the absence of genome editing (Fig. 2 and 3) (5, 3). Figure 2: Gene activation with nuclease-competent...shown to exhibit more limited activity than SpCas9 (2). In many cases this is due both to lower relative...PMID: 24076762. PubMed Central PMCID: PMC3844869. 2. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome...2015): 823-826. PubMed PMID: 26167643. 3. Kiani, Samira, et al. "Cas9 gRNA engineering for genome editing...

...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...were derived from the same source and were incubated on the same plate at the same time. Although the replicate...separate wells (within the same plate) with purified plasmid from the same aliquot/preparation. The replicate...provide the baseline/standard against which other samples can be compared. The Empty Vector Control (mentioned...control in this experiment could be an shRNA—in the same backbone as Plasmid A—that does not target any mammalian...

...human transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In addition, we distribute monoclonal... example through the use of less toxic adjuvants; 2) generating complex antibody chips that allow assessment...assessment of many protein targets in a single sample; 3) producing recombinant, partially humanized rabbit...

...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ...Packaging, and Potency of AAV Vectors. Microorganisms, 12(2), 310. https://doi.org/10.3390/microorganisms1202031038399714...cis-acting elements, which means that they are in the same DNA molecule as the gene that they package. In a.... M., Dobbins, A. L., Li, C., Hirsch, M. L., & Samulski, R. J. (2020). Adeno-Associated Virus Serotype-Specific...

...CcdB.” Biotechniques. 1996 Aug;21(2):320-3. PubMed PMID: 8862819. 2. Bahassi, EM., et al. “F plasmid CcdB... desired insert should not grow.   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials... the ccdB aspect, Gateway takes advantage of the same principle that cells will not propagate while expressing.... While these two strains ultimately perform the same function, there is some evidence that ccdB Survival...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor...individual values match across the board. If the samples or sample conditions change, the multiplexed protocol...validation data, be aware that different machines, samples, and treatments may affect the assay. It’s unusual...

...from the same species and 2) these fragments will be more likely to bind (and block) the same epitopes...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...only antibody option is from the same species as your tissue samples. The bad news: this situation is ...excess secondary with serum from the same species as your primary/sample, and then applying this mixture to...processing and sample preparation can also help reduce the levels of endogenous IgGs in your samples, but know...The culprits are endogenous IgGs in your tissue sample. Most common IHC protocols use two antibodies -... the first antibody. When endogenous IgGs of the same species as your primary antibody are present, your...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...NLuc requires 100-fold less protein to produce the same amount of light from an Fluc or RLuc reaction. In...protein that could be co-imaged with GFP using the same wavelength for excitation. The result was the cyan-excitable... K., et al. (5) imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs...