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Showing: 241 - 260 of 965 results

  1. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...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. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...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...

  3. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...secreted into the periplasmic space to be oxidized and form disulfide bonds (Frenzel et. al., 2013). If you’...applications. An antibody that performs well in immunohistochemistry may not perform well in a western blot. ... than mammalian cells, but mammalian cells can perform human-like post-translational modifications that... If that’s the case, a buffer exchange can be performed using a desalting column or an ultrafiltration...ones, consider increasing the number of washes performed during your harvest. Validation Recombinant antibodies.... This can be due to conformational changes in the antigen or lower detection sensitivity of the assay...S. Sarangapani, Protein aggregation, particle formation, characterization & rheology, Current Opinion ...

  5. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...ultracentrifuge. During the spin, the virions will form a band due to their high density, which can be collected...   Pro tip! An optional media exchange can be performed 12–18 hours post-transfection to help remove toxic...

  6. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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 ...

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

    Type
    Blog Post
    ...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 ...

  9. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...choice, you should perform a functional assay to make sure your protein isn’t forming inclusion bodies ...

  10. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... in spacer acquisition following infection, as a form of “bait” to identify new CRISPR-associated proteins...target binding and cleavage through a series of conformational changes in the Cas13a molecule. Like Cas9, ...

  11. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...changes in gene expression as new body sections are formed. In synthetic biology, FP timers might be able ...isolate particular organelles and their contents, or perform in situ hybridization experiments. In many cases...like the patterning of nascent tissues and the formation of limbs are correlated with changes in gene expression...and developmental timer of neural fiber bundle formation." Journal of Biological Chemistry 276.32 (2001...

  12. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    .... For instance, the fluorophore and quencher may form the base of the stem in an RNA stem-loop structure...and detection - since they frequently undergo conformational changes after target binding, aptamers are ...fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey group selected...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ... 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...

  15. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...and others have reported that FP cysteine residues form inappropriate disulfide bonds in the secretory pathway...modifications including glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized to organelles...fusion protein localization. For example, the formation of inappropriate inter-chain disulfide bonds between...because of the correct folding and fluorophore formation by most, if not all, of the FP fusions. Our moxFPs...

  16. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...Barcoded tubes come in many forms. The 2D barcode can store a lot of information in a small space and is ... treated to easily take in plasmids (transformation). Transformed bacteria are then grown up on antibiotics...received is immediately transformed and stored. Some samples need to be transformed with a micropulser using... (98.2%) transform the first time and can be stored right away. Every week, the lab performs an average...average of 140 transformations. The lab typically transforms newly deposited samples from Monday through Wednesday...delivered by a mail courier and then the journey of transformation and storage begins. Some samples are submitted...barcoded tubes that are scanned into our Laboratory Information Management System (LIMS), a database we use to...

  17. Plasmids 101: Protein tags

    Type
    Blog Post
    ...can also protect against proteolysis. That’s one form of instability. Prokaryotes can also have a hard...Thioredoxin A (TrxA) that improve disulfide bond formation in order to help keep your protein soluble.  ... better assess their function. More detailed information and some protocols can be found in the references...

  18. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ... local family to help, create “fake” family by forming a social circle of people in the same situation... and/or your spouse won’t be at every school performance, chaperone every field trip or cheer at every...
Showing: 241 - 260 of 965 results