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Showing: 221 - 240 of 756 results

  1. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...specify and assemble diverse arrays of neurons, forming the neuronal circuits that will later be refined...traditional cDNA overexpression: isoform-specific gene expression. Gene isoforms refer to mRNAs generated from...untranslated or coding regions. Typically, each gene isoform has a distinct transcriptional start site. Because...transcriptional start sites in order to better understand isoform-specific gene regulation — at least, more straightforward...straightforward than generating cDNA for each gene isoform individually. To highlight this advantage, they...neurons and synapses as well as long-term memory formation. The Bdnf gene possesses 9 distinct transcriptional...with CRISPRa, the Day lab singled out two Bdnf isoforms (I and IV) for investigation. Co-transduction ...

  2. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...vision. Practice drawing and writing everything in a form that is easily visible from the back of the room... vision. Organize your vision into a chalk talk format. Practice. Define your research vision A chalk...find, and how you will interpret and use the information. In contrast, a list of experiments or a laundry...Organize your research vision into the chalk talk format Once you have developed your research vision, you...most of your limited speaking time. Beyond the information you seek to convey in your chalk talk, there'...

  3. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...are required for proper packaging of AAV, but also form secondary structures that are prone to deletion ...and XmaI restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, ...found in the culture media, you should consider performing PEG precipitation. Instead of sonicating the ...

  4. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...antibody-target protein immune complexes are allowed to form. Following incubation, beads are used to immobilize...-40 or triton-X, to preserve native protein conformations and interactions. For difficult to release proteins... in the lysis buffer, keep samples on ice, and perform the IP at 4oC. When choosing the capture antibody... recognizes the target protein in its native conformation. Frequently, the antibodies used for a standard... are completely removed. Wash steps should be performed even with a pre-cleared sample.  Eluting the target...and negative controls for an IP. When possible, perform the IP in parallel with both a sample known to ...order to identify any non-specific interactions, perform parallel IPs with beads only and an isotype control...

  5. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023 under CC-BY 4.0 License.... These variants use different RT enzymes and performed large edits with similar or better efficiency ...potential to reduce negative side effects. This platform offers a promising approach for experimental therapies...we began distribution and has been verified to perform well in standard IHC and western blot protocols...protocols. You can find more specific information on the Wu Lab’s use of Anti-PSD-95 [K28/43R] under the Application...

  6. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ... plates. This prevents satellite colonies from forming while effectively selecting plasmid-containing ...cost-effective, timesaving for transformations. Cons: Prone to satellite colony formation on agar plates, less ...in a plasmid. During the propagation and/or transformation process, some bacteria will either never receive... is nifty because the recovery period after transformation (the dreaded 60 minutes shaking at 37 degrees..., interchangeable with Amp, timesaving for transformations. Con: Expensive.   Kanamycin Kanamycin, or...selection gene. However, it does require a slower transformation recovery period than amp or carb (60 minutes...confers mammalian resistance to G418. Con: Slow transformation   Spectinomycin/streptomycin Spectinomycin...

  7. Getting to Know Expansion Microscopy

    Type
    Blog Post
    Published
    June 20, 2023, 1:15 p.m.
    ...structure. For example, cytoskeletal structures that form continuous lines in pre-expanded samples can sometimes...

  8. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...protein interaction partners (PIPs), the FP fragments form the complete FP because the PIPs they’re tethered...to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent signal...selection of FP fragments and, preferably, some information about the structures of the tested PIPs. Many...fuse the FP fragments. In the absence of this information, you can fuse each FP fragment to the N- and ...your results. Unfortunately, without structural information, constructing appropriate controls may requires...

  9. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...synthetic oscillator, bistable network, stripe pattern-forming incoherent feed-forward loop. Find the plasmids...systems at Addgene! Schematic of the evolution platform to generate TEV protease variants. Catalysis by...the right places, which can provide misleading information on where exactly a structure or protein is located...dissolved at a higher concentration, and results in uniform NB biotinylation. Once induced, the NBs could be...developed fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed...

  10. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...chromatin, the complex of DNA and proteins that forms chromosomes. These long range DNA interactions are... are often detected with chromosome conformation capture (3C) based methods, such as 5C or Hi-C (Han et...generated with current 3C methods provide useful information about chromatin interactions, they are grainy...for answering more questions about chromatin conformations. The Xu lab presented three ways to use CAPTURE... capture of chromatin interactions The Xu lab performed a proof-of-concept experiment to show that CAPTURE...original CAPTURE method. Both CAPTURE methods outperformed other 3D chromatin methods such as ChIA-PET ...is detection of temporal changes in chromatin conformation, like those that occur during development and...

  11. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ...partnered with colleagues at other institutions to form a support network called the MDH CUREs Community...). But is it possible to bring the kinds of transformative educational experiences afforded to those fortunate.... Therefore, he decided to adopt a more focused format, where all students work on malate dehydrogenase...hockey coach all shaped his teaching style, with the former offering excellent training in improvisation and...NADH production.   CUREs are beginning to transform undergraduate education in the sciences by making...

  12. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...location. HDR depends on resection at the break site to form ssDNA and uses the ends of the ssDNA to find a repair...increase HDR frequency by committing a break to performing HR before other factors were loaded onto the ...

  13. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...highlights the dearth of AAV vector information provided in openly accessible forms like publications. Why not ... two years testing and reformulating our AAV construct to optimize performance in our system and yet none...literature, and as a result can leave scientists under-informed with respect to how different serotypes, promoters...this one set of experiments didn’t provide any information on other important factors such as what is the...adjacent areas impacted the experiments we wanted to perform. It required a great deal of time and effort to...packaging viral particles (6), can influence vector performance. Open science initiatives, like Addgene’s Data...negative data and instead focus on getting this information out into the public where it can be utilized ...

  14. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...compaction—on the order of 700 terabytes per gram—and form the X-shaped chromosomes we all know and love. To...enhancers and genome math come in. They help us to transform raw genomic coordinates into meaningful experiments...tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We’ll start with ChIP-seq...

  15. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ... associates with the viral polymerase, which is formed by a complex of two proteins: the large (L) catalytic...measured to quantify (indirectly) transcription performed by the viral polymerase. Alternatively, the mRNA...independently. In addition, experiments can be performed in the absence of some viral proteins (compared... minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...

  16. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ... the anti-sense strand, then the ssDNA template formed will be sense. Therefore, mutagenic oligos need... plasmid. The ssDNA template is generated by transforming a backbone phagemid vector into an E. coli strain...to remove residual starting template. After transformation and harvest, the library is ready for use. ...true if the sense strand is nicked for template formation: template will be anti-sense, primers should be...ease of use, 2) time savings, and 3) ability to perform single site or multi-site saturation mutagenesis...

  17. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    Published
    Feb. 6, 2024, 3:13 p.m.
    ...CD11c-. Finally, B cells are CD11c- and CD123-, forming a population in the lower left(ish) corner.  If...learning how to read a gating strategy.  Other information FACS plots often contain other data. Some will...Abdulaziz University, Kingdom of Saudi Arabia. Bioinformation, 14, 53–59. https://doi.org/10.6026/97320630014053...

  18. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...additional cofactors or using significantly truncated forms that limit its functionality. To make biophysical...C-terminus (or even in the middle of proteins) and performs well in many different antibody-based applications...using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ high rate of spontaneous...indels after DNA repair. B) M. abscessus is first transformed with Cas9 on an integrative plasmid containing...resistance selection marker KanR. This strain is then transformed with the sgRNA cassette on a plasmid expressing...

  19. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ... when bound to DFHBI. It’s also smaller than any form of Spinach - only 49 nucleotides in length - and...aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged ...UTR of mRNA that bind metabolites, leading to conformational changes that control the expression of coding...expression vectors to create a library. Bacteria were transformed, and then sorted by fluorescence-activated cell...

  20. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...causes cells that it infects to fuse together and form syncytias that cause the cells to die. Cytotoxicity... of these review articles to find tables and information on different viral membranes, tropism, and advantages...
Showing: 221 - 240 of 756 results