...Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place in the world...your Addgene plasmid #), and other reagents. F1000Research, among many other journals, has joined a pilot...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...sure your manuscript fits the house style and prerequisites of the journal. Ask colleagues (preferably native... 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...your search and sorting the results (relevance, dates etc). The links for the titles can generally take...you provide, Google Scholar will send you daily updates with relevant literature. Organizing the literature...margins. If you prefer, printing and hand-writing notes is still possible, but not feasible for every paper... University. She is a fan of photosynthesis and writes about autotrophs and biochemistry on her blog. ...Maaike Pols, PhD, a Developmental Editor at F1000Research  ...

...reasons, it is essential to test the selectivity of your antibody. But testing is not one size fits all....important to test for the protocol you intend to use in your experiment. An antibody that demonstrates selectivity...best to determine this by testing them. For intracellular targets, we usually test 3 different fix/perms ...and flow cytometry. F1000Research, 13, 817. https://doi.org/10.12688/f1000research.154034.1 Laflamme, ...synaptotagmin 1 page. The data produced is available on the F1000 gateway and Zenodo community.  Knockdown approach... Correctly interpreting your data therefore necessitates careful design of RNAi sequences, confirmation...like a blood sample containing many different leukocytes. Where data is available from antibody-independent...

...you want to have 100x-1000x coverage of your library: in other words, 100-1000x more reads than number...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds,...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...solid media. The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library...using both pre- and post-amplification DNA and generates PCR sequencing products from both. These products...

...Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen.1000683 Bétermier M, Bertrand P...finishes repair of most types of breaks in tens of minutes – an order of magnitude faster than HDR. NHEJ is...paired end complex The paired end complex then ligates compatible DNA ends together, thus repairing the...Aprataxin, Tdp2) that function to make ends better substrates for ligation. Although we do not describe these...vastly reduces breaks and mutations at off-target sites. Christina Mork contributed to the update of this...

...System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system facilitates the simultaneous... Viruses are intracellular parasites and natural vehicles for genetic information. Therefore they make...Addgene: Follow our viral vector blog posts for updates on the most recent deposits, protocols, and tips...Plasmid Kit Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of...

...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 O’Brien, R. E., Bravo, J...unwinding of dsDNA occur simultaneously, which creates a loop in the target strand that is cut occasionally...

...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on ... clustering and discovery algorithms find new candidates for CRISPR proteins. A type VII is already on...

...and astrocytes Mice No (LY6C1)   Hanlon et al., 2019 AAV-BI-28 CNS: neurons, astrocytes, oligodendrocytes.... It can also effectively transduce astrocytes or oligodendrocytes when paired with appropriate cell type-specific...28 has been tested in C57BL/6J mice and effectively transduces NeuN+ neurons and astrocytes (Huang et al...et al., 2023). Among the capsids tested, AAV9-X1.1 showed the greatest fold change in RNA and DNA levels...Research in Neurobiology, 4, 100086. https://doi.org/10.1016/j.crneur.2023.100086. Challis, R. C., Ravindra... others have done, you’ll need to experimentally test your chosen capsid before using it in an application...elements [promoters and enhancers, miRNA target sites, and recombination target sequences, denoted by ...

...diseases, and several treatments are currently being tested in clinical trials. It is speeding the process ...cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA targeting the...CFTR) by homologous recombination in cultured intestinal stem cells of patients with cystic fibrosis (...also really need to get a handle on how cleavage sites are repaired.” Those insights will be critical for...& Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6. Wu et al. 2020. . “Advances in CRISPR/Cas-based... "Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients...

...STEM graduates and postdocs in academia, industry and other areas of employment? How many graduates and...related to the structure of the biomedical workforce. Quotes taken directly from participants (summarized in...per PI” “Lack of awareness about how the system operates and functions” “Complete lack of information on...of academic research (4). There are currently 17 sites around the USA undertaking BEST programs, including...regulatory, or clinical environment. It remains contested, however, whether a better solution would be to...grassroots organisations that represent postdocs and institutes that employ them, the diversity of trainee backgrounds... and the many different paths that biomedical graduates take in their future careers. To participate in...

...a new legal minimum of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was...blogpost and more recently in a paper published in F1000Research on November 2016. Due to the nature of the postdoc...22nd a preliminary injunction against the FLSA updates was granted nationwide. Again, we at Future of ...getting paid, which we will be analyzing for future updates. But suffice it to say, salaries are showing surprisingly...

...attB sites, the phage integrates into the bacterial genome flanked by two new recombination sites (attL-left...reactions. Recombination of attP and attB sites creates attL and attR sites.   Gateway vectors contain modified...takes place between the attB sites flanking the insert and the attP sites of the donor vector. This reaction...enzyme mix and generates the entry clone containing the DNA of interest flanked by attL sites. As a byproduct...reactions. The BP reaction creates an attL-flanked entry clone. The LR reaction creates an expression clone ... place between the attL sites of the generated entry clone and the attR sites of the destination vector...attB, P, L and R sites that recombine very specifically and directionally: attB1 sites react only with ...

...al., 2002 used an overlapping vector with 440 or 1000bp overlaps to deliver the alkaline phosphatase gene...different splice site donor and acceptor sequences were tested and with the optimal sequences, the group achieved...homologous recombination approach delivers large donor templates using two sequential homology directed repairs...2017) CRISPR-Mediated Integration of Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756...PA (2002) An mRNA Surveillance Mechanism That Eliminates Transcripts Lacking Termination Codons. Science...

...is its speed, labeling neighboring proteins in minutes rather than hours like BioID. APEX has allowed ...and miniTurbo both enable biotin labeling in 10 minutes without any issues in toxicity. TurboID’s signal...give these methods a try.  References:  Branon, Tess C., et al. "Efficient proximity labeling in living...proximity-based labeling methods for interactome mapping. F1000Research, 8.  ...

...research tools. BenchSci collates information about antibodies while FPbase curates information about fluorescent...sometimes formulating new biological concepts. However, rates of reproducibility for preclinical studies, as reported...Project: Cancer Biology have produced similarly low rates of reproducibility. Failed attempts to apply irreproducible... to future work cost both time and money with estimates reaching in the billions of dollars (Freedman ...making it easier to retain searchable, thorough notes on how lab procedures were actually performed, potentially... The publication record will then show that the tested hypothesis is correct (a great success story) even.... Finally there are new publishing outlets like F1000, PeerJ, PLoS, and bioRxiv that make it easier to...

...expected fragments for that plasmid. This phenomenon indicates the presence of a multimer.   Fig. 1:... be used to detect multimers, since restriction sites in multimers usually repeat in tandem along with... of the plasmid monomer (see this article from BiteSize Bio for more information on how different plasmid...Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/elps.201000061. PMID: 20589859. Levene SD. Analysis of DNA topoisomers...

...several labs were able to decipher the journey of lymphocytes from the bone marrow to the thymus where they... tagged with a fluorescent protein or dye. It separates the cells mechanically in a vibrating nozzle, ...How do T cells and/or Neutrophils travel to the sites of infection or damage (Peters et al. 2008, Chtanova.... 2008, Kamenyeva et al. 2015)? How do blood monocytes patrol blood vessels during immune responses (...Salomonnson et al. 2012). Finally, it is worthwhile to test the photostability of your fluorescent protein when...flank fluorescent proteins in a plasmid with loxP sites such that their expression is turned on or off in...with Toxoplasma gondii." PLoS Pathog 5.7 (2009): e1000505. PubMed PMID: 19578440. PubMed Central PMCID: ...