We narrowed to 844 results for: tat

...type of experiment is called a protein fragmentation complementation assay (PCA). In the Split-BioID method...FRET is another example of a protein fragmentation complementation assay that can be used to study binary...individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID, a spatiotemporally controllable...Protein Interactions. Split-BioID is a protein complementation assay where BirA*, an E. coli biotin ligase...proteins. One caveat of Split-BioID is that the orientation of the BirA* fusion matters! Schopp et al saw...interact with the bait proteins. Another potential limitation of Split-BioID is its speed of labeling. BioID...

..., or for generating mutated sequences for site-directed mutagenesis.  Quantitative polymerase chain reaction...genotyping. It’s extremely useful but is not as quantitative as other methods of PCR. Theoretically scientists...reaction, qPCR Quantitative polymerase chain reaction (qPCR) also known as real time PCR is a PCR technique...material in your sample.  qPCR can also be used to quantitate RNA levels using reverse transcription-qPCR (...sequences that are rare, for example, rare alleles or mutations. ddPCR also does not require a reference or standard...is determined and then analyzed using Poisson statistics to determine the concentration of the original...PCRs that use a single pair of primers, like those stated above. Multiplex PCR is also used in high throughput...

...heterogeneity dictates clonal responses to the acquisition of leukemic driver mutations. Cell Stem Cell... during differentiation after they introduced mutations that drive leukemia (Figure 3). Figure...Figure 3: Using LARRYv2 libraries for state-fate analysis in ex vivo HSC expansion cultures. A) Experimental...Experimental design. B) UMAP of cell states in ex vivo expansion cultures from days 7, 14, and 27. Reproduced...useful for studying the impacts of other cancer mutations and non-genetic diversity in human cancer stem...binds to AMPA receptor subunit GluA1 found at excitatory synapses (Rivera & Huang, et al., 2025). AF was...determined cells. This version of ATLAS works only at excitatory synapses; however, its modular design allows ...

...contains a mutation or modification, this should be included in the plasmid name. Mutations are generally...abbreviation of the mutated amino acid currently at that position. In contrast, unmutated or Wild Type (wt...your plasmid name. This is often a 3-6 letter representation of the gene (or DNA sequence). pBACKBONE-hGene...change. The proper way to denote an amino acid mutation is to list the one letter abbreviation of the ...   Example: If the Glutamine at position 295 was mutated to an Alanine, Q295A.  Mutant version: pBACKBONE-Flag-hGene...

...Considering citations to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles...to 1975 and is based on the average number of citations for recent articles. Of course, a publication ...measure scientific impact. Even if we stick to citation counts for a moment, it’s clear that articles ...articles in non-elite journals went from 27% of all citations in 1995 to 47% in 2013 – yes, almost half.   The...Accessibility is surely also behind the increase in citations for older publications. (As an interesting aside...the amount of protein in a solution.) But are citations and journal publications really the best way to...

...COVID-19 cases each week. Statistical Modeling Note -- If statistical modeling isn’t up your alley,... estimated total of 380,000 deaths worldwide (statistics provided by Google on 6/4/20). Many countries... open-ended question is difficult to answer quantitatively without broad, unbiased data. Therefore, we...order several plasmids at once, breaking the statistical independence assumption of the Poisson process...process. This modification also eases the interpretation of the parameters we infer, since scientific productivity...remind the reader that this is not a causal interpretation -- the effect would generally not be the same...His research uses applied machine learning and statistical methods for fundamental scientific discovery ...

...parents looking for experts to come give guest presentations. If you’re a scientist thinking about reaching...already discussed the idea of my giving a CRISPR presentation to their students. I didn’t think of this opportunity...postdocs, and other working scientists, creating a presentation for 8th graders was a little intimidating. Too...received from the teachers, I formulated the presentation as follows (which could be adapted for any topic...do? I spent more time on this section of the presentation because, in my opinion, it was more important...teachers with whom I worked, for feedback on my presentation, she said “it was such a valuable extension ... of CRISPR. Two of the examples I used in my presentation had only become public in the previous two weeks...

... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) because any engineered DNA-binding...region of the transcription factor IRF-1. Immunoprecipitation, using an antibody targeting the 3xFLAG tag...purification of a specific genomic region, numerous adaptations of this system have been made in different fields...also has great potential for improvements and adaptations that can provide an increased understanding of... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR." Biochemical and biophysical... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171...and Hodaka Fujii. "Insertional chromatin immunoprecipitation: a method for isolating specific genomic ...

... an invitation to join the Advisory Board of Addgene, I accepted the offer without any hesitation. I hope...transcription by using locus-specific chromatin immunoprecipitation (locus-specific ChIP). These methods consist...consist of insertional chromatin immunoprecipitation (iChIP) and engineered DNA-binding molecule-mediated...molecule-mediated chromatin immunoprecipitation (enChIP), both developed in the lab. In June 2014, Dr. Fujii joined ..., we developed the insertional chromatin immunoprecipitation (ChIP) (iChIP) technology, which uses locus-tagging... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP). Fujita T, Asano Y, Ohtsuka J, Takada... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Fujita T, Fujii H....

...differentiated somatic cell reverts back to a pluripotent state, which then proliferates and is redifferentiated...differentiated cell type to an undifferentiated state and then redifferentiation into the cell type of...Conversion of adult somatic cells into the pluripotent state. Redifferentiation - Conversion of the pluripotent...Dedifferentiation to an intermediate pluripotent state is not always obligatory in cell conversion processes...back to their most primitive pluripotent stem cell state, through transdifferentiation adult somatic cells...factors) More Chances of accumulation of harmful mutations More, due to selection from more cycles of proliferation...objective is to fix a disease-inducing genetic mutation in a patient, trying to transdifferentiate any...

...such a small package. But don’t let the simple presentation deceive you; pooled libraries can be quite challenging...verified through NGS sequencing to confirm library representation. Transfection of the DNA pooled library and...transduction of cells, resulting in cell population mutated by the viral pooled library. These cells can then...protocol that amplifies without disrupting plasmid representation. A good place to start is with a protocol for...library by next-generation sequencing (NGS). The representation of your amplified library should be very similar...page, you can find protocols for sequencing, representative NGS results (read counts or another visualization...library and verify that you have not skewed the representation of your pooled library during the amplification...

... an enzymatically “active” state rather than reverting to an inactive state like Cas9 or Cpf1. As a result...proteins like SaCas9 and Cpf1 can circumvent the limitations associated with SpCas9. Cas13a (previously referred... HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a molecule results in...system, Cas9 uses both a tracrRNA and crRNA to facilitate binding and cleavage of target DNA respectively...molecule through a uracil-rich stem loop and facilitates target binding and cleavage through a series ... to activate programmed cell death or a dormant state for bacterial cells that have been infected with...Zika virus, genotype human DNA, identify tumor mutations within cell-free genomic DNA, and detect COVID...

....1c02201. LiLac sensor for lactate Stop and smell the flowers - or rather, the lactate! The latest biosensor...for lactate developed using a high-throughput multiparameter screen (Koveal et al. 2022). Lactate is a...previous lactate biosensors, and its affinity is better-matched to cellular concentrations of lactate. The...ZAP70 (ZtSH2) which binds only the phosphorylated state of an immunoreceptor tyrosinase-based activation...around 70 mM and 100 mM, respectively. Introducing mutations and using potassium-binding protein homologues...

..., I promise it isn’t.) The intuitive, qualitative interpretation of this gel is misleading. We can see...causes misleading qualitative conclusions, and it’s also problematic for computational quantification. ...experimental parameters. This is a job for a semi-quantitative western blot. Most western blots benefit from...Loading controls are even more important for semi-quantitative western blots. No pipetter is perfect, and proteins...lysate (10–40 µg) probed with β-actin (1:2000). Representative of three independent blots. B–C) Graphs showing...high-abundance single-protein controls in semi-quantitative immunoblotting. Journal of Neuroscience Methods...

...the trust you’ve put in us when you choose this status for your plasmids. In fact, almost half of all ...routinely review deposits that are left in the hold status and reach out to depositors about them, but we ...ensure that plasmids don’t get left behind in this status — a win-win situation for all. Addgene will release... a quick email at [email protected] with the citation and your deposit or plasmid ID numbers and we’...’ll be happy to update the plasmids to add the citation and release the hold. Increasing impact Making...before publication and we can update the plasmids’ citation when the paper comes out later. Or, if a project... these goals. And we can update your plasmids’ citation to the final journal article version any time ...

...bactericidal) or slow growth/prevent cell division (bacteriostatic); however, the distinction between the two ...categories may be a bit of a gray area as some bacteriostatic reagents can kill bacteria when used at high...subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin B polypeptide...inhibits protein synthesis (elongation step) Bacteriostatic 10 ug/mL Chloramphenicol   Binds 50S ribosomal...ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes.     ...inactivates the gene and acts as a marker for the mutation. When designing these types of experiments it ...not to use the same resistance cassette for the mutation and for plasmid selection. Additionally, scientists...

...DNA with phi29 polymerase prior to fragmentation can greatly facilitate downstream manipulations. Phi29 ...activity without the need for genome sequencing or annotation (5). In the event that genome sequencing can ...to enzymatic digestion, however, a physical fragmentation strategy is preferable. Nebulization involves...associated with cloning large fragments. After fragmentation, repair sheared ends with T4 DNA polymerase ...gently down onto a plate of colonies. Mark the orientation of each membrane on its plate before moving on...your membrane to several freeze/thaw cycles to facilitate cell lysis: alternating between 5 min at -20°...sequence the inserted DNA and characterize your new putative host cell-binding proteins! Many thanks to our...

...professionals has made possible a significant qualitative and quantitative expansion in the 2015-2016 academic ...does research at UK in the same field that I do: prostate cancer. She has guided me in my next steps. For...baseline self-assessment survey to determine their expectations of the program and their baseline skills. Mentors... as mentors to Spanish mentees, mentors also facilitated the development of bidirectional USA-Europe professional... pre-program self-assessment survey defining expectations of the program and their baseline skills and...improvements in writing fellowships, giving oral presentations, and preparing for job interviews, expanded ...Spanish scientific community overseas. Official representatives from different Spanish universities and high...

...others.  Meditate. Meditating during the day is a great way to take a mental break, while meditating right...Organize your data and prepare publications or your dissertation Organize data that you already collected into...into figures for papers or presentations. Learn a new skill for analyzing and visualizing your data, such...needing to have collected data. Work on your dissertation, even if your defense isn’t for a while. Getting...Data Hub to help the research community. Set expectations for your productivity Set a routine and stick...

...  An MTA is simply a contract or agreement that states the terms of use for a tangible research material...MTAs.  Addgene does a lot to ensure that we can facilitate a fast and agile MTA process. This is important...deposits in our repository. From 2004 to 2023, we facilitated 784,795 MTAs with 8,798 organizations in 111 ...111 countries. From 2019 to 2023, we averaged facilitating 89,477 MTAs per year.  The MTA Process at Addgene...MTAs for plasmids in our repository. Instead, we facilitate the signing of MTAs by the Provider and Recipient...signature on any subsequent MTA for that plasmid facilitated by Addgene — basically, instead of having to ...shared reagents should be used. Setting clear expectations from the beginning makes the sharing process...