We narrowed to 1,015 results for: NTS;

...errors in the design of cloning strategies set experiments back days or even weeks. Primer design was done...principles. For every task, we envision what the user wants to do and make the path to accomplishing their goals...is an ongoing process that involves iterative refinements in response to customer feedback. An example ... and display frequently used genes and control elements. Development of this tool required creating a ...variant of a feature. Instead, we identified common variants, and then crafted a detection algorithm that tolerates...sequence has changed. Identifying plasmid control elements Coding sequence features are relatively straightforward...straightforward to define, but for control elements such as promoters and transcription terminators, the boundaries...

... the field of grants. What is a grant professional?  In the past 5-10 years, the Grants Profession has...collaborations, and commitments to and proof of compliance with the grantor’s requirements as well as state... for grants, the amount of grant funds available and the increasing regulatory process for grants has ...grant consultants who offer grant training, mentoring and coaching services, including Smartegrants, to ...choice exam.  The bottom line on being a grants professional The Grants Profession is in a rapid growth phase... Inc., a national grant consulting firm, and Smartegrants, a grant professional development firm The path...established grant professionals is learning to succeed in grants through trial and error, through winning and losing...

.... Now is the time to make bold statements. While some of the statements may be slightly far reaching, ...consider the reviewer's comments. Whether the proposal is funded or not, the comments help gauge room for ...government and academic institutions. New graduate students may assume that their advisor alone applies for... allot some if not all grant duties to trusted students. For example, if the proposal call aligns with...write a grant!”, “I don’t have time for this if he wants data by next week!”, and “This is his job!” filled...various skill building opportunities that graduate students and postdocs should actively pursue. Proposal ...italic, or underline font formats helps these statements stand out from the rest of the text. Specific...

...single 96-well plate for HDR that satisfy all requirements. To alleviate this bottleneck, we began building...whole or in part. For example, you can analyze experiments designed outside CrispyCrunch, or you can input...step of mutating guide sequences in a way that prevents re-cutting but does not disrupt gene expression...that worked with NGS data were: Crispresso CrispRVariants AmpliCan We chose to adapt Crispresso by Luca...re-create a match for the active guide. With these constraints in mind, we implemented the following algorithm...Design donor RNA Design primers Review and order reagents Perform your wet lab experiment Analyze the data...high-throughput design and analysis of CRISPR experiments. It has certainly boosted our productivity in...

...levels and affect behavior. A fact to consider: Rodents can hear ultrasonic frequencies (> 20 kHz, produced...

...around the globe, we’re now sharing over 100,000 reagents including plasmids, ready-to-use viral preps, ...

...(i.e. the plasmid complements the deletion). If the phenotype is corrected, students can be relatively... multiple enzymatic steps (5). In their experiments, students test whether or not proteins from the fission...extensive rearrangements in the S. cerevisiae lineage (7). In our class research project, students transform...in other experiments (Figure 2).   Complementation reveals conservation of function Students have used...Boston College. Students in the course work in teams of three in sections of 15 students. Within each section...also used to teach students about restriction mapping and western blotting. Students who participate in...authentic research experiences to undergraduate students (1, 2), but educators face significant challenges...

.... Both awardees emphasized the contribution of students in their labs and the importance of the strong... what can be done with this technology to help patients. Dr. Stoffels and others in the audience discussed...technologies. Addgene is proud to be helping get CRISPR reagents out to academic labs. It is our hope that robust...

... cloning efficiency measurements. We performed additional efficiency measurements using similarly sized...number of transformants); the greater the number of inserts, the lower the number of transformants. To compensate... Table 1). These improvements (5- to 12-fold increases in the number of transformants while maintaining...reaction volumes were 20 μl (1, 12 fragments) or 25 μl (24 fragments). The future of assembly DNA assembly...Assembly is commonly used to assemble 2–10 DNA fragments in a single “one-pot” reaction to form complex... site is no longer present, allowing multiple fragments to be assembled in an ordered, pre-determined ...background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion of the...

...CRISPR applications in plants.     Transgenic yeast as source of dietary supplements Dr. Mrinalini Nair,...well as developing yeast as source for dietary supplements! Development of a bacterial expression system..., bacteria, and fungi that reside within living plants and that produce compounds with various antimicrobial...More Optogenetic Tools Like pDawn  Making cassava plants resistant to cassava mosaic virus Dr. Fiona Robertson...make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy DNA viruses, similar to...interested in applying gene technology to bacteria and plants. Her research involves cloning cry genes from the...against local insect pests. By developing biological agents such as insecticidal proteins, the use of chemical...

...responses in patients. They were able to tweak this assay to study SARS-CoV-2 spike variants. “We created...circulating variants. This allowed us to create pseudoviruses that represent lineages and variants that are...we’ve seen the emergence of several SARS-CoV-2 variants coincide with increased vaccination efforts. So...effective are the current vaccines against the variants? Alejandro Balazs, a Group Leader at the Ragon... of current vaccines differed depending on the variants (Garcia-Beltran et al., 2021). Antibodies from...Moderna or Pfizer vaccine could neutralize many variants, including the B.1.1.7 (Alpha variant), B1.1.298...well as they could for the wild-type. However, variants such as the P.1 (Gamma variant) and the B.1.351...

...share these improvements? You could publish a paper that happens to include your improvements, or maybe ...the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem many...information, you’ll have the option to add additional documents. By uploading your protocols onto your plasmid... to your protocols when they begin their own experiments. Need to submit an updated protocol? Just email...them public for others to use, and add your own comments. Currently you can find protocols from NEB, various...storage and inventory, managing orders, tracking students and research projects, electronic lab notebooks...trials and pricing options for academic labs, departments, industry, and more to fit every lab's needs....

...Quantifying the abundance of thousands of protein variants simultaneously  With increasing sequencing power...individual’s genome. But when they find genetic variants, what does that mean? Previous research shows ...approximately 75% of all protein-coding pathogenic human variants are decreased in protein abundance, possibly due... due to misfolding. But measuring variants one at a time is time consuming and unscalable.  Melissa Chiasson...Washington sought to address the effects of genetic variants in a high throughput manner. She helped develop...a method that measures the effects of sequence variants on abundance simultaneously called VAMP-seq, for...protein variant library is monitored by fusing the variants to eGFP followed by an internal ribosomal entry...

... Taq buffer with MgCl 2 1 μl dNTP mix (10 mM each nt) 2.5 μL Forward Primer (10 μM stock) 2.5 μL Reverse...total + 10% (3.5μl) = 38.5μl dNTP mix (10 mM each nt): 1µl x 7 reactions = 7µl total + 10% (0.7µl) = 7.7µl...thousands of identical copies using a simple set of reagents and a basic heating and cooling (denaturing and...of the DNA segment to be amplified. When these reagents are combined in an appropriate buffer, a series... the newly synthesized strands. Materials List Reagents for each 50µL PCR: Thin-walled PCR tube Ice Bucket... on ice Set up a 50 μL reaction (Keep all your reagents on ice): 2 μL Template DNA (10 ng-500 ng) 5 μl...

...Plasma membrane (Plants) PIP2a mCherry Maria Harrison 61206 pCMU-PDESr Plasmodesmata (Plants) PDLP1 mCherry... made up of microtubules, actin filaments, and intermediate filaments. These structures give the cell ...GFP-cortactin Actin filaments Cortactin EGFP Anna Huttenlocher 32907 pEGFP-rNFM Intermediate filaments (neuronal...Intermediate filaments (neuronal) Nefm EGFP Anthony Brown 11908 pEGFP-N1 alpha-actinin 1 Actin filaments alpha-Actinin...Actin filaments Cortactin mCherry Christien Merrifield 36201 pLifeAct-mTurquoise2 Actin filaments LifeAct...pLifeAct_mScarlet_N1 Actin filaments LifeAct mScarlet Dorus Gadella 85056 pLifeAct_mScarlet-i_N1 Actin filaments LifeAct ... Actin filaments ITPKA mTurquoise2 Dorus Gadella 58473 pEGFP-C1 F-tractin-EGFP Actin filaments F-tractin...

...tools that were specifically designed for use in plants. These resources include tools for research using...useful toolkits and plasmid collections for use in plants. The tables below highlight plasmids and plasmid...empty backbones to express your gene of interest in plants. You can search the table for features you are ...for plant transformation, including regulatory elements, coding sequences, basic CRISPR parts, and parts... of fusion constructs in transgenic analysis of plants. Return to top Genome Editing The following table...of our most popular plasmids for gene editing in plants. Please refer to Addgene's Plant CRISPR Resources...selection of plasmids expressing different functional variants of Cas9. ID Plasmid Description PI Genome Editing...

... 20-nucleotide (nt) guide sequence that is complementary to the DNA target. The 20-nt guide region binds...the 20-nt Guide Sequence Identify a TAM site (5′-TTGAT-3′) in your target gene. Choose a 20-nt sequence...bacterial selection. The process for cloning the 20-nt guide RNA is identical for both vectors. Here, we...Dra2TnpB) to create a genome editing system for plants (Karmakar et al., 2024). This version of TnpB is...a different TAM (5′-TTAT-3′). The unique TAM requirements expand the number of genomic sites that can ...have a familiar targeting mechanism. Small RNA elements called right-end RNA (reRNA) or omega RNA (ωRNA...genome editing is the efficient delivery of editing reagents into cells. Viral vectors, because of their natural...

...antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments - Fab fragments are created... called the antigen binding fragments (Fab). The tips of the Fab fragments contain highly variable regions...Affinity Reagents There are several alternatives to antibodies, ranging from antibody fragments (shown ...Structure Isotypes Antigen Specificity Other Affinity Reagents Handling Visualization Signal Amplification Application...Multiplex Assays Selection Validation Affinity reagents are molecular tools used in the lab to specifically...foreign bodies, are the most commonly used affinity reagents. Antibodies are Y-shaped proteins, naturally produced...interest is introduced to the animal, which then mounts an immune response to it. Once the immune response...

...multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements, and other Cas...scale edits by integrating segments of DNA flanked by specific repeat elements into the genome. CRISPR-associated...approach. Engineered CRISPR systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated...mechanisms of CRISPR and some of the ways the CRISPR components are leveraged to achieve specific goals, such...the Cas enzyme. Basic Mechanisms of CRISPR The requirements for a CRISPR knockout assay are simple. You’...mechanism. Multiplex Genome Engineering Many CRISPR experiments involve editing more than one gene. In these ...RuvC and HNH, can be altered to reduce cleavage events to a single cut (nick) or no cuts. Cas9 nickase...

...safety feature of lentiviral vectors is that the components necessary to produce an infectious viral particle...Vector Guide for more information about lentiviral components, generations, viral production, and viral safety...expression. See Ventura et al. (2004) for more variants. Tyler Jacks 19319 pLJM1-EGFP 3rd CMV-driven EGFP...transgene or shRNA. See Szulc et al. (2006) for more variants. Patrick Aebischer and Didier Trono 11795 pLL3.7... 3rd Modification of pLL3.7 with added genetic elements known to prevent epigenetic silencing. Expresses...markers. See Campeau et al. (2009) for additional variants. Eric Campeau 39481 pLenti-puro 3rd CMV-driven...selection. See Shin et al. (2006) for additional variants. Iain Fraser 15948 pLOVE 3rd Tet inducible gateway...