We narrowed to 963 results for: tes
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TypeBlog Post...below). Plating the transformation on ampicillin plates containing X-gal allows you to identify cells that...that when a portion of the gene is deleted, it creates a non-functional đť›˝-galactosidase. Complementing...this deleted portion, for example on a plasmid, creates a functional đť›˝-galactosidase. Blue-white screens...
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Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2
TypeBlog Post...can rapidly and efficiently evaluate Cas9 cutting sites and activity (Knudsen et al., 2018). To create ...lab compared editing at three different genomic sites in the AAVS1 locus using GEmCherry2. In this experiment...again that sgRNA structure and genomic recognition sites play a large role in CRISPR-Cas9 efficiency. They... -
Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors
TypeBlog Post...information, making it available to everyone, that creates more breakthrough and innovation. With any kind...Suffolk University Law School. Suffolk University graduates on average 20 PhD’s a year and we have a very ...Committee at the Greater Boston Food Bank and participates locally in Read To A Child's "Lunchtime Reading... -
Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification
TypeBlog Post... start, the team inserted AsLOV2 into conserved sites in all eight surface-exposed loops of an mCherry...SOS), a guanine nucleotide exchange factor that activates Ras. They expressed this OptoNB-SOS fusion protein...membrane-localized mCherry. With darkness, SOS binds Ras and initiates the extracellular signaling-regulated kinase (... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog Post...together. I also prefer the number system to using dates as labels, since I often work on multiple projects...sure that the original code remains in tact! Take notes Comment in your scripts: Comments on what a given...who are still learning the basics. However, these notes take up a lot of space and don’t tell you why you... -
3 Challenges in Plant Synthetic Biology
TypeBlog Post...infancy. A long timeline from transformation to testing and a lack of well-characterized genetic tools ...excited to see how this exchange of genetic tools facilitates plant science. Challenge #3: Intellectual property...in 2004, she accepted a commission in the United States Air Force. She left active duty to pursue a degree... -
Promoters
TypeGuide...the other hand, the process in eukaryotes is much more complex. Eukaryotes require a minimum of seven transcription...actually begins. Only eukaryotes and archaea, however, contain this TATA box. Most prokaryotes contain a sequence...are common in prokayotes, specifically bacteria, but have also been discovered in eukaryotes. Operons consist...binding is very different in bacteria compared to eukaryotes. In bacteria, the core RNA polymerase requires...Transcription is tightly controlled in both bacteria and eukaryotes. Promoters are controlled by various DNA regulatory...promoter also contains transcription factor binding sites, but mostly contains regulatory elements. Eukaryotic...Specific Drosophila promoter containing Gal4 binding sites Bacterial Promoters Promoters in bacteria contain... -
Summer SciComm: Preprints
TypeBlog Post...Photo courtesy of Henry MĂĽhlpfordt, under a CC BY-SA license. Many scientists and advocates have argued...manuscript is drafted. Preprints were one of the key routes for researchers to share information about the ... -
Antibodies 101: Affinity Reagents
TypeBlog Post...large number of targets, and can require specific conjugates. And, of course, once you have them, you’ll need... along with a higher likelihood of aggregation (Bates, 2019). They are small enough to be produced in ...delivery systems. Elsevier Science & Technology Books. Bates A, Power CA. David vs. Goliath: The Structure, Function... -
Antibodies 101: Selecting the Right Antibody
TypeBlog Post...options. With limited time and budget you cannot test them all, so how do you know which ones to try? ...immune complexes are smaller than a primary:secondaries:conjugates complex. If you are trying to visualize.... The 5 pillars include genetic strategies that test expression in knock-out or knock-down down tissues... -
What's New in CRISPR - November 2021
TypeBlog Post...with the desired resolution of prime editing intermediates. Building upon the PE2 and PE3 systems of their...introduce epitope tags and RNA transcription start sites. The lab further demonstrated that PRIME-del can...RT), which induces two double-strand breaks and deletes the intervening sequence. As with other prime editing... -
Choosing the B(right)est Fluorescent Protein: Photostability
TypeBlog Post... oxygen concentration may affect photobleaching rates (Shaner et al, 2008). This brings us to the key ...application (Goedhart et al, 2012). The third issue relates to the environment of the fluorescent protein. ...The issues with quantification of photobleaching rates can be summarized as follows: Instead of determining... -
CRISPRainbow and Genome Visualization
TypeBlog Post... cross-talk with the other orthologs. Ma et al. tested their dCas9 variants using gRNAs specific for telomeric...individual colors to 6. A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a...separate vector. Figure 2: CRISPRainbow generates 6 unique colors and a white composite. gRNAs with... -
Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages
TypeBlog Post...a new legal minimum of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was...22nd a preliminary injunction against the FLSA updates was granted nationwide. Again, we at Future of ...getting paid, which we will be analyzing for future updates. But suffice it to say, salaries are showing surprisingly... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog Post...CASANOVA responds to blue light quickly. Within minutes of inactivating the CASANOVA complex, fluorescent...genes-of-interest (GOI) into two independent expression cassettes on a single plasmid (Grefen and Blatt, 2012). ...proteins that bind to either the light- and dark- states of the photoswitchable cPYP, which changes in conformation... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog Post...caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle progression and enhances tumor... TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate... 2004). In fact, Sanford H. Barsky and his team tested the influence myoepithelial cells had on the conversion... -
Seven Tips for Using LinkedIn as a Scientist
TypeBlog Post...keep track of how you met the connection, as these notes are saved in the messaging stream. Unfortunately...LinkedIn is not the platform for politics, family updates, or click bait of any kind. For more on inappropriate...However, do comment on other people’s news and updates. Share your own good ideas, events and favorite... -
Troubleshooting Your Plasmid Cloning Experiment
TypeBlog Post...antibiotic in your plate might not work. Check the plates! Streak on them an E. coli  strain known to be ...growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic. Many small colonies... different temperature (30°C) for the growth on plates and in liquid culture. If you don’t get any positive... -
Working with Nuclear Receptors
TypeBlog Post...and disease states, their circadian regulation, as well as in important physiological states such as adipogenesis...therapeutic indices, including hypertension, cancer, diabetes, cardiovascular disease, cholesterol gallstone... -
Adeno-associated virus (AAV) Guide
TypeGuide...available inside of the small AAV capsid and directly correlates to the size of the DNA removed from the viral... transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...is crucial for efficient AAV replication as it activates the necessary cellular machinery to produce large...Random integration of rAAV genomes in non-homologous sites of the host genome can occur, although at a low ...a slash (e.g. AAV2/5), where the first number indicates the serotype of the genome and the second number...specific cell types of interest has been to create and test capsid libraries with random mutations or insertions... PNS (e.g. AAV-PHP.S) in rodents and non-human primates. Read our blog post on systemic capsids for more...