We narrowed to 723 results for: tat

...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...

... cutting edge genomic technologies to map the mutations that are present, and use prior knowledge (data...One central bottleneck is that most recurrently mutated cancer genes are rare and most of the individual...characterized? Some have developed exciting computational algorithms, such as SIFT or PolyPhen to approach...well as reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic...High-throughput Phenotyping of Lung Cancer Somatic Mutations. Berger AH, Brooks AN, Wu X, et al. Cancer Cell...perform additional functional assays for these mutations and map other cellular functions and phenotypes...to build saturating libraries of all possible mutations in many major cancer genes. These should be used...

...track of computation work comes with new sets of challenges. One challenge with computational analyses... of biologists are finding themselves using computational approaches to analyze large data sets (myself...what works, and what does not work. Careful documentation will keep you on task and will prevent you from...especially since you’ll want to dig in to the computation, but they will serve you well as you perform ... the analyses: Goal: Before you begin, briefly state the goal of the analysis, including background on... you to perform this analysis. Having a clearly stated, specific goal for each analysis will help you ...

... homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids used...microorganisms, yeast have both a stable haploid and diploid state which is useful for genetic analysis, as well as...apparently independent of a known specific sequence) dictate the replication of these vectors. S. pombe plasmids...missing nutrient. Scientists can exploit these host mutations by including a copy of a functional gene which...This can complement ura4- S. pombe, but the complementation is weak. yes   LYS2 L-lysine yes - Grow on... S. pombe  no no *FOA in fission yeast induces mutation in the ura5+ gene in addition to ura4+. leu1...

... DIO (Double-floxed Inverse Orientation) or DO (Double-floxed Orientation)) is a powerful tool that provides...channelrhodopsins that, in turn, trigger neural excitation, whereas yellow light activates halorhodopsins...resulting fusion gene is inserted in the antisense orientation with respect to the promoter to prevent its expression...recognition sites (e.g., loxP and lox511) in opposite orientations. Since Cre does not cause recombination between...therefore locking the opsin into the correct orientation to be transcribed (Figure 2) (Sharma and Zhu,... arousal in mice previously subjected to steady-state general anesthesia (Taylor et al., 2016). These ...strains of Halobacterium halobium differing in pigmentation." Biochemical and biophysical research communications...

... (e.g. accessibility due to chromatin status). No computational prediction is ever perfect, but this can...cell to use an alternative ATG downstream of the annotated start codon. Likewise, we avoid target sites that...fluorescent tag or the introduction of a specific mutation, one generally relies on homology directed repair...exact location of the TSS. Different databases annotate the TSS in different ways, and it has been shown...experimental conditions used, led to no detectable mutations (Veres et al., 2014). When working with single-cell... effective, so long as care is taken in the interpretation of results by requiring multiple sequences ...Musunuru K (2014) Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human...

...employed to mutate whole antibody sequences, degenerate primers can be designed to selectively mutate specific...research tools like western blots, ELISAs, immunoprecipitation, and flow cytometry. The better these tools...improve an antibody, but don’t have a specific mutation in mind. Generating a random library of mutants...structural framework then guides the specific mutations you choose, whether it's tweaking the antibody-antigen...and increase the chances of finding beneficial mutations. In other words, structure-guided evolution allows...cell surface display are common. ELISA enables quantitative measurement of antigen-antibody interactions..., yeast, or mammalian display, allow for the presentation of your mutant libraries on the surface of cells...

...: Summary of Rosella’s excitation and emission spectra. Component Excitation Emission pH Range DsRed.T3... inhibitor. While this method does provide a quantitative measure of autophagy, it only measures bulk ...final read out of this assay. This is also a quantitative measure of autophagy, but, like the protein ...DsRed.T3. See Table 1 for a summary of Rosella’s excitation and emission spectra. DsRed is the permanent ... 508 nm ~6.5 - 9 *Optimal sequential excitations wavelengths as determined by Rosado et al: see... T., & Klionsky, D. J. (2008). Chapter 3 The Quantitative Pho8Δ60 Assay of Nonspecific Autophagy. Methods...

...assays include immunoprecipitation mass spectrometry (IP/MS) and chromatin immunoprecipitation (ChIP). In ...In IP/MS, a target protein is first immunoprecipitated - along with its associated protein complexes - ...tag” such that these tagged proteins can be immunoprecipitated using a standard tag-specific antibody (Jarvik...double-strand breaks (DSBs). DSB repair by HDR facilitates in-frame insertion of tag-encoding sequences ...efficiency across different stem cell types. This has facilitated easy generation of clonal knock-in lines that... design, remember to incorporate PAM-blocking mutations (e.g. convert the NGG PAM to NGC in your repair...2018), however, if you have any questions, do not hesitate to contact me at [email protected]. Keep ...

...and immunology. They can be used for qualitative and quantitative measurement of protein expression in...protein via fluorescence or chemiluminescence, precipitate the protein out of solution, or isolate cells...epitope of the target protein. However, small mutations in the DNA of the hybridoma can occur, known as...types or treatment conditions. They can be used quantitatively if you also load the gel with appropriate controls...media from cultured cells. ELISA can be used to quantitatively measure the amount of protein present. The ...ELISA (Enzyme-linked Immunosorbent Assay). Immunoprecipitation Sometimes, instead of just detecting the ...immunofluorescence), or by a colorimetric reaction facilitated by an enzymatic reaction. Figure 5: Immunofluorescence...

...and methylated states. Thus, this system is a reliable tool in reporting methylation status of these sites...the headlines, with sotires like the potential devastation of the palm oil industry through epigenetic effects...grandchildren; in this study,epigenetic changes were statistically significantly associated with reduced life-span...syndrome is characterized by low muscle tone, short stature, cognitive disabilities, problem behaviors, and...epigenetics; techniques include chromatin immunoprecipitation, fluorescent in-situ hybridization, methylation-sensitive...methyllysine binding domain causes a structural change (facilitated by the flexible linker) that brings the fluorescent...

...2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA cleavage upon co-expression and spontaneous...has been cumbersome. To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors...or invert a specific chunk of DNA. However, a limitation to this strategy is that each "bit" of information...This leads to 2048 (211) possible combinations of states. Each of the integrases is supplied on its own ...more powerful LID, the authors recently used computational protein design, phage display and high throughput...system. How does it work? RUSH is an ingenious two-state assay based on the reversible interaction between...synthase pair is encoded on a single plasmid and facilitates the incorporation of a variety of bio-orthogonal...

...but unsure how best to prepare. Maybe you’re a presentation veteran, but have difficulty answering questions...what you would like to say to help build your presentation. For example, when thinking about discussing... Practice, practice, practice: Rehearse your presentation with people inside and outside your field. Your...and PI can provide feedback on your technical presentation, but those outside of your field can point out...you a chance to determine how to tailor your presentation so that it can be understood by your audience...It’s likely you won’t know every answer – don’t hesitate to be honest and tell your audience just that....attendees can get sucked in to thirty-minute-long presentations, which constrains the number of posters you ...

...the late 1980s, scientists even created a codon adaptation index (CAI) that is based on the codon usage ...codon bias. The idea is that scientists can make mutations throughout a gene of interest based on an organisms...) to name a few. This could affect synonymous mutations which have even been linked to some diseases. ...human genes contain a region where a synonymous mutation can be harmful (Sauna and Kimchi-Sarfaty, 2011...tabulated from international DNA sequence databases: status for the year 2000. Nucleic Acids Research 28:292... Understanding the contribution of synonymous mutations to human disease. Nat Rev Genet 12:683–691. https...10.1038/nrg3051 Sharp PM, Li W-H (1987) The codon adaptation index-a measure of directional synonymous codon...

...single protein, computationally described as a fusion of four Cas7 proteins and a putative Cas11-like protein...patient's specific mutations. With a drag and drop approach, however, the specific mutations wouldn’t matter...evolution, were also working on the project. Using computational observations, Koonin and Makarova found that...which can be caused by thousands of different mutations, all found in the same gene. If we only have small...matter – the mutated copy could be replaced by simply dragging and dropping in a functional copy of the entire...