We narrowed to 42 results for: ins/1000

...produces point mutations at a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate.... The Moffat laboratory maintains a website for the TKO library that contains up-to-date information and...(3) but its portability to other bacterial strains remains seriously limited, as prior optimizations are...ligase to biotinylate proximate proteins in the cell. These candidate proteins can be enriched via biotin ...of fusion proteins, requires less biotin, and shows enhanced labeling of proximate proteins. Overall BioID2...optimization of industrial yeast strains. However, industrial yeast strains are more difficult to genetically...which “bait” proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with...

...Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place in the world...journal. An increasing number of funding bodies insist that the research data resulting from their funding...your Addgene plasmid #), and other reagents. F1000Research, among many other journals, has joined a pilot...many journals use to verify whether a manuscript contains all the necessary information to be of maximal...results more valuable. Some journals, including F1000Research, also ask for the raw data behind the figures...several rounds of revision before acceptance. F1000Research uses a novel form of peer review that occurs... Maaike Pols is a Developmental Editor for F1000Research, responsible for assessing the scientific content...

... organization issues – F1000 just announced the launch of their F1000Workspace.  Give your favorite searching...though you may need to be on a network with an institutional subscription), archived on PubMed Central, or...Check out the hashtag #365papers on Twitter for inspiration and accountability. There are a few different...reprints and papers with scribbled notations in the margins. If you prefer, printing and hand-writing notes...guest blogger! Johnna Roose is a biochemistry instructor in the Department of Biological Sciences at Louisiana...Maaike Pols, PhD, a Developmental Editor at F1000Research  ...

...sector throughout their careers (McDowell et al, F1000Research, 2014; Dolan, F100Research, 2016). These events...students and postdocs broadly – and thus PI’s and institutions in turn. The first is the passing and upcoming...the NIH recommended pay scale – used by many institutions to set postdoc wages. The second event of note...simply being able to categorize all postdocs at institutions, will be challenging. At the same time, there...level, FoR is currently compiling a database of institutional responses to the FLSA ruling. Similarly, from...Research: a perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...from the Future of Research Chicago Symposium F1000Research, 2016. Kyle T. Dolan, Joseph F. Pierre and Erin...

...and flow cytometry. F1000Research, 13, 817. https://doi.org/10.12688/f1000research.154034.1 Laflamme, ...approach, which focuses on validating antibodies against markers of the human blood leukocyte populations...cases, we would recommend a knockdown approach instead. In this approach, you’re looking for an antibody...synaptotagmin 1 page. The data produced is available on the F1000 gateway and Zenodo community.  Knockdown approach...permanent knockdown, consider using shRNA vectors instead. With both siRNA and shRNA, it can be difficult...induce/suppress the expression of many related proteins, not just your protein of interest, which can ...interest, which can be difficult to find for many proteins. In practice, it was necessary for us to combine...

...fluorescent proteins. Additional fluorescent protein tools Photoactivable fluorescent proteins (proteins that... Blue-green fluorescent proteins like EGFP Yellow-orange fluorescent proteins like TagRFP, tdTomato, DsRed...danger. In this post we’ll focus on how fluorescent proteins can and have been used to monitor cellular movements...specimen. It allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated...that have been tagged with different fluorescent proteins or dyes. 3. Two-photon microscopy Two-photon excitation...Cellular trafficking studies enabled by fluorescent proteins The optimization of GFP and its derivatives by...changed everything. For the first time, fluorescent proteins gave researchers the ability to track immune cells...

...) if NHEJ joins the distal ends together. Similarly, it may be possible to direct insertion of an exogenous...Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen.1000683 Bétermier M, Bertrand P...of breaks) by NHEJ-dependent repair (“pop-in” insertion) provided a template containing compatible overhangs...Renouf B, Renaud J-B, Sallmyr A, Ruis B, Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin M, ...

...you want to have 100x-1000x coverage of your library: in other words, 100-1000x more reads than number... the DNA prep is larger. And since the library contains many plasmids, care must be taken to ensure that...plasmids in the library are identical except for the insert; there’s no need to sequence other parts of the...

..., and Cas11. Type I-E Cascades contains all five of these Cas proteins, while type I-C Cascades do not...boundaries defined by anti-CRISPR proteins or running into other proteins naturally bound to the DNA. Cas3...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 O’Brien, R. E., Bravo, J...The helicase domain is then activated, and Cas3 begins unraveling the double-stranded DNA (dsDNA) in what...Cas9 vs. The Other Cas(s) CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems INTEGRATE: Bacterial...

...Kit Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of 96 retroviral... for packaging, however, the isoforms of these proteins, as well as the viral LTRs differ. Therefore, ...Multiple Lentiviral Expression System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system...

...model organism, they insert into the host’s genome. Then, the transposon insertion sites are detected with...transposase proteins (TnsA, TnsB, TnsC), and 4) the donor DNA or mini-transposon that contains the DNA cargo... out of the laboratory. F1000Res 9:135. https://doi.org/10.12688/f1000research.21018.1 Munoz-Lopez M, ...and result in human disease. While transposon insertions can be detrimental, it also drives genomic evolution...that moves around. This DNA sequence encodes the proteins the transposon needs for transposition. Transposition...transposition.  Target site: Different transposons insert at different DNA sequences or target sites. Integration...duplication of this target site sequence at the site of insertion.  Types of transposons Transposable elements ...

...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on ...and understanding of CRISPR, and perhaps provide insights and ideas for your own research! Classification...bacteria and nearly 100% in archaea, though Class 1 remains largely underutilized compared to Class 2. Class...effector complex but are defined by the accessory Cas proteins in the Cascade complex. Despite a tenuous grasp... genes, Cas1 and Cas2, which are critical for inserting short fragments of foreign DNA into the CRISPR...CRISPR in more depth, the Jennifer Doudna lab maintains an encyclopedia of Class 2 CRISPR systems called...a CRISPR-associated transposase (CAST) that can insert large fragments of DNA. The V-F variants, which...

...Research in Neurobiology, 4, 100086. https://doi.org/10.1016/j.crneur.2023.100086. Challis, R. C., Ravindra...exists to prevent substances in the blood, like proteins and viruses, from crossing into the brain. While...increase the chances of inducing an immune response against the capsids or transgenes. Sensory neurons and ...as the type of transgene delivered: fluorescent proteins, optogenetic sensors, CRISPR elements, etc... ...capsids express differently in different mouse strains, a phenomenon that we are calling “strain-specific... but is more effective than AAV9 in many other strains, including FVB/NCrl, 129S1/SvImJ, DBA/2J, and C57BL...table from Huang et al, 2019 for affected mouse strains). However, other capsids cross the BBB via interactions...

... Alzheimer's disease & Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6. Wu et al. 2020. . “Advances...in monoallelic diseases In 2013, a team led by Jinsong Li from the Chinese Academy of Sciences found that...independent team led by Hans Clevers at Hubrecht Institute in The Netherlands used the CRISPR/Cas9 genome...handle on how cleavage sites are repaired.” Those insights will be critical for scientists’ ability to influence...addition, the current work in developing anti-CRISPR proteins aid the limitation of these mutations, to control...

...were at institutions focused only on tracking hours, and 22% of postdocs were at institutions where no... of some public institutions) to cancel plans to raise salaries. What are institutions doing after the...planning unionization at their institutions. One clear message that institutions have sent, with the lack of...their institution is doing from Future of Research over social media, not from their institution), is ...raise were: postdocs at institutions like Stanford and the Whitehead Institute, where salaries had already...street from these institutions) are raising salaries. Postdocs at the Salk Institute in La Jolla California...This post discusses how a nationwide injunction against the FLSA is affecting universities' decisions to...

... to train our young scientists for futures both inside and outside of academic research (4). There are...industry, regulatory, or clinical environment. It remains contested, however, whether a better solution would...University, has recently published an article arguing against reducing the number of graduate students accepted...grassroots organisations that represent postdocs and institutes that employ them, the diversity of trainee backgrounds...Research: a perspective from junior scientists." F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...

...limited to traditional restriction enzyme cloning. Instead, you can choose a molecular cloning technique that...takes place between the attB sites flanking the insert and the attP sites of the donor vector. This reaction... inhibits the growth of CcdB sensitive E. coli strains, most colonies should contain the desired, recombined...vector.   Method B: TOPO-cloning of the desired insert into an attL-entry-TOPO vector. TOPO cloning adds...Method B to create an entry clone: TOPO cloning the insert into an attL-containing entry vector.   Method...interest and a attL-entry vector. This fragment is inserted in a multiple cloning site (MCS) of an attL-containing... create an entry clone: Restriction cloning the inset into an attL-containing entry vector.   Pro...

...within living cells. By coupling fluorophores to proteins, pioneers of this technique were able to directly...FRET-based biosensors, and interactions between DNA and proteins. FRET, in theory, is an easy technique, but it...to studying an unknown interaction between two proteins, these parameters are difficult to overcome but...FRET_teaching_module.pdf Day RN, Davidson MW. Fluorescent proteins for FRET microscopy: monitoring protein interactions...Bioessays. 2012 May; 34(5):341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance...

...interactions as well as be used for both soluble and insoluble proteins, which can be a challenge for classic antibody...can take hours to label proteins and APEX2, although it can rapidly label proteins, requires H2O2 which ...interacting proteins in one simple experiment. PCA is an in vivo technique where two proteins are fused ...interaction of two proteins while also biotin labeling and purifying additional proteins from a complex of... and an RNA component that contains an RNA motif of interest. Any proteins that bind the RNA motif will...interacting cell surface proteins with a biotin tagged LPETG peptide. Biotin tagged proteins can then be purified...There are approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells...

...al., 2002 used an overlapping vector with 440 or 1000bp overlaps to deliver the alkaline phosphatase gene...transgene.The AAV encoding part A of the transgene contains a promoter and the 5’-fragment of the transgene...donor. The AAV encoding part B of the transgene contains a splice site acceptor followed by the 3’-end ...methods. The AAV encoding part A of the transgene contains the promoter, the 5’ half of the transgene, a ...homology. The AAV encoding part B of the transgene contains the region of homology, a splice acceptor sequence...help prevent the accumulation of such truncated proteins. Examples of use:  Ghosh et al., 2008 used a...directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to the site in the genome...