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Showing: 401 - 410 of 997 results
  1. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems, these...nucleotide sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes...targeting constructs. In the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the ... the cells that have randomly integrated all or large portions of the plasmid. Deliver your construct ...insert GFP into the genome, we’d create a plasmid similar to the one shown below, where the sequence of GFP...CRISPR is very good at making knockouts, knocking in large sections of DNA can be more difficult. Addgene depositors...
  2. Plasmids 101: Optimizing Plasmid Yields

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    Blog Post
    ... might need to use multiple techniques for a particularly stubborn prep. Check the bacterial strain Increase...strains, including HB101 and derivatives, release large amounts of carbohydrates that can inhibit the cell...levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression...choice of strain will ultimately depend on the particular features of your plasmid, it is advisable to ...coli strains. Increase the volume Starting with a larger culture volume can improve plasmid yield. Our lab...gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around 220 RPM,...
  3. Anatomy of a Plasmid Page at Addgene

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    Blog Post
    ...depositing lab. There may be other plasmids with similar names in the collection, but you can use this name...some of Addgene’s plasmids is an indication of popularity. A yellow flame indicates that a plasmid has ... on our website, one of the available filters popularity (e.g. only showing those plasmids which have ... than 100 times).     I guess it's cool to be popular, but how do I know if a plasmid is actually useful...If the depositor has indicated that there is a particular mutation found within the gene/insert, this information...means you can see which labs have already used a particular plasmid, and which experimental systems it was...
  4. Genome-wide Screening Using CRISPR

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    Blog Post
    ...role of individual genes or entire pathways in a particular disease process. While CRISPR is certainly not... mutant cells to identify genes involved in a particular phenotype. CRISPR can be readily scaled up for...fluorescence-based cell sorting and screened for a particular phenotype. For example, mutant cells can be used...cell types that are of low abundance are not particularly well suited for genome-wide screening. Avoid...targeting the same gene can be strong evidence that a particular gene is actually important for a given phenotype...Lentiviral CRISPR Libraries. Current Protocols in Molecular Biology 115: . https://doi.org/10.1002/cpmb.8 ...
  5. Tips for Using BLAST to Verify Plasmids

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    Blog Post
    ...results. The default setting is “Optimize for Highly similar sequences (megablast)”, which is very fast and ...often unreliable, we suggest using the “Somewhat similar sequences (blastn)” program for blastn so that ...your sequencing trace file. Unlike megablast, the regular blastn program uses a smaller word size and lower...protein sequences (nr)” database as it contains the largest number of protein sequences. Just below the BLAST...options and for our suggested blastx customization. Similar to nucleotide sequences, proteins often have repeated...COBALT for aligning multiple protein sequences, particularly for comparing different isoforms. In addition...
  6. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

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    Blog Post
    ...transistor-like digital genetic switches that enable cellular computing. These logic gates built from transcriptors...of promises to not assert property rights for particular engineered genetic sequences; but it’s only the...computer languages are free to use. Of the 15 most popular computer programming languages, 12 out of 15 are...are now available exist within the context of the larger bioeconomy, and that it’s been 40 years since the... on the domestic market generates $300 billion dollars every year. That's two percent of the U.S. gross... we anticipate plugging into the probiotic and cellular therapeutic space. If you were to look forward...
  7. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

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    Blog Post
    ...information while using less resources and are particularly useful when trying to screen a limited number... libraries have been shown to outperform other popular CRISPR libraries (see below). Let’s take a look...Brunello library was more effective than other popular CRISPR libraries in the depletion of essential ...discriminate between non-essential and essential genes similarly to the Brunello knock-out library while mitigating...between 150-75 nucleotides upstream of the TSS. Similar to the Brunello and Dolcetto library, sgRNAs were...about the advantages of pooled CRISPR libraries for large-scale functional screens Read this blog post about...
  8. Getting to Know Expansion Microscopy

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    Blog Post
    ... Boyden Lab, ExM has been steadily growing in popularity with an abundance of protocols and examples of...not something you find in the average cell and molecular biology lab, and EM is limited in the number of...provides new insights into the cytoskeleton of malaria parasites including the conservation of a conoid..., Mohar B, Eddison M, et al (2022) Visualizing cellular and tissue ultrastructure using Ten-fold Robust... Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures using expansion microscopy (U-...Wijesekara P, et al (2023) Magnify is a universal molecular anchoring strategy for expansion microscopy. Nat...
  9. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...Interestingly, he also adds to the disproportionately large number of honorees to have trained under a previous...fortunate enough to apprentice with top scientists to larger groups of trainees at an earlier point in their...protein with easily measured activity, MDH is particularly amenable to in-class labs. Provost ultimately... at the American Society for Biochemistry and Molecular Biology Annual Meeting. Some of my students published...and green fluorescent protein. Biochemistry and Molecular Biology Education. 38(5):317-323. Knutson, K.,...dehydrogenase as a research model. Biochemistry and Molecular Biology Education. 38(5):324-329. Kowalski, J....
  10. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...specializes in developing novel technologies to analyze molecular mechanisms of genome functions such as epigenetic...transcription and epigenetic regulation. To elucidate molecular mechanisms of regulation of genome functions, ...isolate specific genomic regions while retaining molecular interactions. For this purpose, we developed the...merit of depositing plasmids to Addgene has been spectacular. We've got more than one hundred requests, and...been very difficult for us to respond to such a large number of requests by ourselves without Addgene.... experiences in research labs. What are some similarities and differences you have observed as you have...
Showing: 401 - 410 of 997 results