We narrowed to 353 results for: plat
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TypeBlog Post...samples through our lab using barcoded tubes and plates. The critical pieces of information for a given...100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then ... database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for forward...shipped along with their corresponding glycerol plate on dry ice to the sequencing provider. Each sequencing...
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Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post...genes. For this you would need to provide a DNA template to repair the double strand break via homology...Cas9/sgRNA and one vector for your specific DNA template. Using this two vectors system, Manuel Gonçalves...This technology therefore represents a potential platform that could be realized via simple switching of...tools can also be used to deliver Cas9 and DNA templates in somatic cells of adult animals in order to ... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known... -
The AAVantages of AAV in CRISPR Screens
TypeBlog Post... need to deliver the Cas protein altogether. Platforming AAV Interest in interrogating and mapping the...glioblastoma, and generate programmable screening platforms for mouse tissues. Your AAVancement could be next...highly scalable cell type-specificin vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https..., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. J. (2023). Transcriptional linkage analysis ... -
Plasmids 101: Knockout/Knock-In Plasmids
TypeBlog Post...homologous recombination using a repair template. These templates can include loxP sites to create conditional...recombination. The concept is simple: deliver a template with a modified version of the targeted sequence...sequence to the cell which will recombine the template with the endogenous gene. Here, we'll describe the ... -
Tips for arabidopsis transformation
TypeBlog Post... from an LB plate. In my hands, a 5 mL starter culture inoculated from a colony on a plate will reach ..., check out Harrison et al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase...difficult. Don’t leave your plants on selective plates for too long: even resistant plants will be affected... -
Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments
TypeBlog Post... to copy and modify templates (e.g. http://openwetware.org/wiki/Protocols/Template). Nature Protocol Exchange... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial... -
CRISPR-mediated Plant Base Editors
TypeBlog Post... base editing platforms for plant systems. Figure 2: Plant base editor platforms and their molecular... They can, but they require other base editor platforms. Hua et al. constructed a number of plant base...editing experiments in plants Choose a base editing platform depending on the location of your target base ... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog Post...strategy for increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs ...homologous recombination approach delivers large donor templates using two sequential homology directed repairs... the gRNA target site that mediated the donor template’s integration so that the site is reconstituted...which results in the integration of a larger donor template. Drawbacks: Transgene expression varies with... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog Post...broth and on the agar plates. Mycoplasma colonies are easily visible on the agar plates. This method is capable...detection, DNA staining, fluorescent tagging, agar plating, etc. Be wise to choose any of the techniques listed... (Uphoff and Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard’ for detecting... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog Post...in 16 wells of a 96-well plate, allowing 6 conditions to be screened per plate. The results indicated that... pools were prepared consisting of one target (template) strand and four upstream probes and four down-stream...strand covering all 16 possible NN pairs in the template. The probes were designed such that each possible... -
Addgene is Stopping Use of X (formerly Twitter)
TypeBlog Post...LinkedIn. We are also exploring other social media platforms with active scientific communities and welcome...functionality and features that previously made the platform a mainstay of conversation in the scientific community...considering a gradual move off X as a response to major platform changes, such as limiting access and increased... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...cloning have been utilized to create recombination template vectors, to perform multiple site mutagenesis ...Engineering TALENs containing variable-repeats using Platinum Gate system Interested in optimizing TALEN assembly... single-strand annealing (SSA) assay. This new Platinum Gate TALEN Kit utilizes a 4-module assembly system... plasmids (ex. p1HD, p1NG, p1NI & p1NN) in the Platinum Gate system contains an identical variable repeat...reports. For additional information on using the Platinum Gate TALEN Kit, please see the Yamamoto lab’s ... -
Save Time and Money by Making Your Own Competent Cells
TypeBlog Post...incubation step and go straight to plating if your transformed cells are being plated on agar with ampicillin selection...competent cells, don’t forget to do a negative control (plate your cells without adding plasmid) to make sure ... -
Fluorescent Proteins 101: Fluorescent Biosensors
TypeBlog Post...Method Notes Quantification of a process Plate-based assays (plate reader) Detect the average amount of ...fluorescence across a population of cells (i.e. in a plate well). Use to track a process (e.g. enzyme activity... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog Post...enough time to completely digest the template DNA. Note—templates must have been isolated from methylation...sensitive restriction enzyme DpnI to remove the templates. Digestion products are then transformed directly... -
Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools
TypeBlog Post...Biology 2018 Pubmed PMID 29379139. The “Riboglow” platform for visualizing RNA production and localization...cobalamin binding riboswitches used in the Riboglow platform at Addgene. Braselmann E, et al. Nature Chemical...a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer a ... -
Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools
TypeBlog Post...fields of biology and enable the conversion of RNA templates to DNA but suffer from several important shortcomings...probability of the polymerase continuing to copy the template rather than dissociating). MarathonRT, isolated...structured HCV genome (~9.6 kb of highly structured template!). MarathonRT also faithfully transcribes RNA ... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog Post...repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9...worms have laid eggs, hygromycin is added to the plates to select for candidate knock-ins. These candidate... -
KLF Research Plasmids
TypeCollection...Keisuke Kaji Jonathan Katz Nathan Lawson Kathrin Plath Miguel Ramalho-Santos Derrick Rossi Michael Ruppert...