...methylase mutation (GATC) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes... mutation (CCWGG) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F-mcrA Δ(mrr-hsdRMS-mcrBC...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F- gyrA462 endA1 glnV44 Δ...maintenance of a plasmids with R6Kγ origin; contains a mutant allele of the pir gene that maintains the plasmids...screening and routine cloning, nalidixic acid resistant. endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44...plasmids, ligated DNA, and libraries, nalidixic acid resistant.  endA1 glnV44 recA1 thi-1 gyrA96 relA1 lac Hte...

...relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said...comprehensive understanding of how cancers become resistant, an emerging theme is that they do so by activating...signaling pathways involved in drug resistance Wood wanted to find out which particular pathways drive resistance...pathways, they identified a set of one to three mutant complementary DNAs (cDNAs) representing core nodes...of - and that’s thrilling to us,” Wood says. “We want to make these available as widely as possible.” ...of signaling pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014...

...annotate genetic perturbations that may play significant roles in cancer pathogenesis. Their work culminated... any mutations that that confer proliferation advantages will be revealed in the absence of growth factor...mutations were found to be “neutral,” having no significant effect on cell viability compared to a wild-type...Approximately ~22% of the mutations, however, significantly increased cell viability, and these “activators...passenger mutations from the “active” drivers is important for the development of targeted therapeutic interventions...dozens of wild-type genes and their associated mutants in other model systems. It is possible that a given...

...using Twitter to find phages to treat an antibiotic resistant bacterial infection, she shared that tweet...for phage may not be needed if other antibiotics work. “We don’t want to cry wolf to the phage community...woman with CF whose infection has failed all #antibiotics. We need lytic non-lysogenic phage URGENTLY to... a repository of phage data and information. “We want to be the go-to place for phage information because...

...(IPI), in which Addgene will distribute recombinant antibodies (rAbs) developed at IPI. The first IPI ...excited to launch our Antibody Data Hub, an open-access repository for antibody validation data. We’ll...world. That included 2008 pooled libraries, 290 antibodies, 18,359 viral items, and 166,247 plasmids. We... our distributor partners, with the launch of antibodies in China and expansion of viral services available...

...scale: Look at the mEmerald variants first, followed by the color variants. The mEmerald plasmids offer...Davidson lab prepares to shutter its doors. “We wanted to give all of these vectors to the scientific ... FSU. Mike is really the one who had been very adamant that we distribute to as many researchers as we...reagents. "We are proud to be able to bring such an important collection to the scientific community," says ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

...ever been the target of an ‘-omics’ study or is resistant to specific compounds, links to publication references...cross-references with cell line catalogs and collections. Importantly, Cellosaurus also identifies problematic cell... the most common contaminator line.  For those wanting to access Cellosaurus programatically, their API... helpful information on the known features and relevant literature, or to decide which line is best for...scientists all over globe, with some surprising, and important, findings being shared. For instance, recently...around! If you’re excited about this resource and want to learn more, check out the Cellosaurus FAQ! References...

... have pushed the boundaries in neuroscience by granting targeted control over neuronal activity. While..., both techniques offer researchers different advantages and limitations.  The four main factors in which...studies, however, require a permanent intracranial implant to be surgically installed to allow for light delivery... delivery. Chemogenetics poses the advantage of being less invasive and more flexible as it relies on ...Invasiveness Mice require permanent intracranial implant Noninvasive Controlling Stimulation ExogenousEasily...controlled EndogenousNot easily controlled Unique advantages Mapping the brainDissecting circuits Studying...

...direct, specific and sensitive detection of the plant hormones strigolactones By Fabienne Furt Current... detect strigolactones (SLs) are costly, non quantitative or require heavy optimization. Claudia Vickers...fluorescence intensity, it provides a direct and quantitative detection of SLs. The authors also showed that... known to trigger seed germination of parasitic plants, this approach was also used to build and validate...Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and reduced...

...ve realized my figures are just as important, if not more important, than my writing. Initially I didn...design software, like Adobe Illustrator, and I didn’t want to pay for these programs either. But with a little...presentations, manuscripts, or social media. If you want to draw your own figures Google Drawings Google ...through your university or your public library. If you want to use pre-drawn images in your figures Biorender...New icons can be requested, although there’s no guarantee Biorender will create it. Additionally, you can...

... our ReFigures on the therapeutic use of anti-CD47 antibodies in immuno-oncology. Funding agencies such... Cataloging a reagent’s performance as in this antibody ReFigure Analyzing replications as in this ReFigure... ReFigure to coherently save analyses with the relevant figures from a literature review. No more hunting... and good for science ReFigures are published instantly and are editable by the author at any time. Insights...preprints, data, software, micropublications) in grant proposals. ReFigures are one such research product...

... both of these cases, transposons combine the advantages of viruses and naked DNA while eliminating some...therapy - avoiding the use of viruses is also important for social and regulatory reasons. 1.) The transposon...Zsuzsanna Izsvak’s lab screened many SB tranposase variants for hyperactivity in mammalian cells. Their efforts...transposase (as determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In...inducible expression vectors were developed into 16 variants with different selection markers (blasticidin,...as a nonviral vector for CD19-directed chimeric antigen receptor (CAR) therapy. SB is a valuable tool and...

...expression. Experimental output is measured and quantified. Created with BioRender.com.   Optogenetic...2020). Light-gated ion channels are useful when you want to activate or inhibit specific neurons without ...pathway. Photoswitches are great for teasing out important branch points along a signaling cascade that may... activated. Created with BioRender.com.   Important considerations for using optogenetic tools Once...optogenetics or biosensors in one system. There can be substantial overlap in the activation curves (Marshel et ... fidelity with the stimulus. For example, if you want to stimulate at a fast frequency without missing...can wreak havoc on your experiments, so it is important to understand the details of how each tool performs...

...administered systemically, without causing disease? Recombinant adeno-associated viral vectors, or rAAVs, can...serotypes can be useful to evade neutralizing antibodies developed from prior natural exposure or rAAV...route of administration (Wu, Asokan, et al., 2006). Want to know if a capsid is a good option for your experiment...61779-370-7_6 Tabebordbar, M., Lagerborg, K. A., Stanton, A., King, E. M., Ye, S., Tellez, L., Krunnfusz.... Directed evolution of a family of AAV capsid variants enabling potent muscle-directed gene delivery ...., Gerfen, C. R., Schiller, J., Dudman, J. T., Hantman, A. W., Looger, L. L., Schaffer, D. V., & Karpova...Karpova, A. Y. (2016). A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons. ...

...findings can be predictably applied to future work. Importantly, no one should expect that every study should...who have complaints about the unreliability of antibodies. IPI is working to address this issue directly...directly by creating well characterised antibodies against all human extracellular proteins with community-...research tools. BenchSci collates information about antibodies while FPbase curates information about fluorescent...and use computer based analysis techniques, an important component of reproducibility is making sure that...figures. Cause #2: Hidden “negative” data We all want to be able to tell a good story when presenting ...journal’s perspective, probably not what most readers want to read. As Susanna Bachle points out in her talk...

...reporter-based) each come with their advantages and disadvantages. Sanger sequencing is simple and the...1B below). Alternatively, if an attB-attP pair is anti-aligned (sites facing opposite directions) on a ...from multiple primer pairs, one can infer the predominant state of registers within a sample using a metric...multiple registers. However, it is only semi-quantitative. It won’t tell you the distribution of states...states in a sample; instead, it will suggest the predominant state of registers in a sample based on a similarity...of inputs (see Figure S9 of our paper (1)). We anticipate that, depending on the context and application...

...reporter can also quantify Cas9 cleavage, thus exhibiting its versatile utility in quantifying multiple gene...versatile in vivo fluorescent reporter for rapid quantification of base-editing and Cas9 cleavage activity ...several cell types is the lack of a rapid and quantitative editing assay. To help accelerate the expansion...plasmid that allows for rapid fluorescence-based quantification of base editors that utilize dCas9 fused to...frame-shift mutation within the mCherry reporter. Quantification of editing efficiencies simply involves tracking...

...restriction sites are present in a given sequence. You want to choose enzymes that: Do not cut within your...Next, we need to examine the DNA sequence that we want to amplify and design primers that will bind to ...primer design: Because we are cloning an ORF, we want to clone from the start codon (ATG) to the stop ...reaction Run PCR to amplify your insert DNA. It is important to use a high fidelity polymerase to minimize .... The fidelity of the polymerase becomes more important the longer the expected PCR product is. You should...used. No matter which polymerase you use, it is important that you sequence the final product. Additional...

...treated a tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102,...with EcoRI and selected for E. coli transformants that were resistant to both. When this proved successful...generic term for any extrachromsomal hereditary determinant”. His proposal, however, was basically ignored...was the tool of choice for molecular biologists wanting to study bacterial genetics. This all changed thanks...

...physiologically-relevant systems, to catch all the important interactions. It’s also important to determine...speak, with collaborators in Paris and New York. We want YOU to join the fight against coronavirus We are... a lot of background in our study - so you might want to be careful with these clones (and help us figure...been quite the administrative headache receiving constant email requests and tracking their fulfillment ...