We narrowed to 855 results for: NTS

...link together multiple fragments, allowing ordered assembly of multiple components into a destination vector...suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!   Why use multiplexed...(here, O1-4) adjacent to the cut sites “link” fragments together and drive ordered assembly into a Cas9...and thus the highest frequency of genome editing events. Yamamoto Lab Multiplex CRISPR/Cas9 Assembly Kit... with the Gersbach lab plasmids, multiple Cas9 variants are available: wt humanized Cas9, D10A nickase...clone your own entry vectors containing other Cas9 variants to use with the MuLE system. Multiplexing from...are also available from the Yang lab for use in plants. The mammalian multiplex systems use the Csy4 RNA...

...can vary "Lab assistants" usually wash glassware, prep media, etc. "Research assistants" are more involved... researchers, as well as graduate students (your teaching assistants), about their experiences. Research...investigator (PI) Runs the lab Writes a lot of grants Varying degrees of engagement with what happens...degree "Rotation student" - some PhD programs have students rotate through a few labs, ~10 weeks per lab, ...way to answer those questions. I’ve met graduate students who enjoy the thrill of acquiring a piece of knowledge...with your labmates and mentors Many of the grad students and postdocs I’ve met have done things that I ...about careers in science are geared towards grad students and postdocs—people who are a little ahead along...

...assembly, assembling fragments in multiple steps to avoid using multiple fragments that share homology ...cloning method, but it is limited by its sequence constraints. The long overhangs must not contain the dNTP...ligation-independent cloning (SLIC), eliminated many of LIC’s constraints by relying on the power of homologous recombination...the linearized vector by T4 digestion; then the components are combined and annealed. The resulting recombination...up a new world of multicomponent assembly, as fragments with overlapping sequence homology can be assembled...of multicomponent assembly with SLIC. Multiple fragments are PCR amplified to add 5′ and 3′ homology regions...vector. 5′ overhangs are generated on all the fragments and the vector, specifying the order of assembly...

...few Plasmids 101 posts, we focused mainly on the elements required for plasmid maintenance within an E. ...different cell types and each one requires different elements for vector propagation. This post, along with ...and explain how they differ from the bacterial elements previously described. Why do scientists use yeast...complex internal cell structures similar to those of plants and animals. Unlike bacteria, yeast can post-translationally...in the lab typically contain all the necessary components to allow shuttling between E. coli and yeast ...the bacterial cell. Similarly, the specific ORI elements included within a yeast vector determine how the...replicated and maintained within the yeast cell. These elements control not only the number of plasmids found ...

...functions. Experiments come next, testing the correctness of the hypothesis. The results of experiments can ...already encouraged by “Broader Impacts” requirements for certain grants) (28). As one young scientist, Matthew...Politics and science are intrinsically linked: governments exist in reality and science is how we learn ... to protect their rights (2). Less formally, governments are the organizations that make and enforce the...information about reality informing people and their governments on what outcomes are actually possible for a ...informed fashion. For science to be useful to governments, however, inconvenient data must not be ignored...climate change page on whitehouse.gov, freezing grants, and proposing to defund aspects of research at...

... the Rare Genomics Institute to provide plasmid grants to researchers working on rare diseases through...group recently reviewed and reported that PMM2-CDG patients suffer from immunological defects. These are characterized...almost every aspect of immunology, participating in events such as cell development, recognition and migration...migration, the immune compromise observed in CDG patients is not surprising. However, the molecular and cellular...cellular mechanisms behind the immune defects in CDG patients are still poorly studied and understood.  In line... to lower the burden of immune symptoms on CDG patients and to increase their quality of life by reducing... the lab will be greatly facilitated by CRISPR reagents and other specific gene sequences available from...

...the FP(11) and FP(1–10) fragments. When Protein A and B interact, the FP fragments can assemble the full...al. 2017 under CC-BY license. Many split-FP fragments are named according to the number of strands in...10 and 11 to yield large (1–10) and small (11) fragments. In several cases an FP(1–10) fragment may be ...be compatible with multiple FP(11) fragments, even from a different base FP (for example, EBFP2(1–10) can...Venus Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis...pBiFC-VC155 Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis...Chang-Deng Hu mVenus pET-BiFC contains both fragments of split mVenus (aa 155), includes I152L in N-...

...PAM with no sequence constraints, or at least has looser or different requirements. Luckily, there are ...most gene targeting experiments. Between SpRY and several of the other Cas variants described, nearly all...need for gene editing in research, the old PAM constraints just won’t cut it anymore. Say hello to PAM-free...PAM must be selected. When it comes to CRISPR experiments, therefore, you often must make a choice about...region will do the job. What matters for these experiments is cutting efficiency so that the greatest effect...that don’t utilize NGG   There are plenty of Cas9 variants which utilize alternative PAMs. These alternative... generating Cas enzymes with alternative PAM requirements (Zhao et. al, 2023). xCas9 has been shown to...

...CRISPR Affinity Purification in situ of Regulatory Elements) method. The original CAPTURE method was published...CRISPR Affinity Purification in situ of Regulatory Elements: The original CAPTURE method The original CAPTURE...of a stable cell line that co-expresses three components: dCas9 with an added biotin acceptor site BirA...genomic location of interest  When these three components are co-expressed, the gRNA targets the dCas9 ...then developed by detecting these chimeric DNA fragments by PCR or next-generation sequencing. Figure...diagram). Cross-links are reversed so that DNA fragments can be detected by PCR or deep sequencing. ...number of cells (~5x107). Together, these two requirements prevent the use of CAPTURE with primary cells...

...organizing events. I contacted local professional organizations and nonprofits. I volunteered at events like...resources daily to promote CINSO events and recruit speakers and participants for our professional development...Science Outreach, or CINSO. We organize fun science events for adults and professional development opportunities...best I began viewing my activities as types of experiments. I searched for new opportunities with an open...wasn’t something I liked – well, I was used to experiments not working, so just like in lab, I sucked it... – call it “troubleshooting.” Trying out new experiments and new ideas is nothing new to a scientist. ... opportunities provided by your programs and departments Graduate school was an excellent time to try ...