... meeting on the calendar even if it is just 15 minutes a week. Clearly identify constraints and boundaries...only way and may not even be the best way. This facilitates success and trust. Micromanaging is bad for everyone...Update frequently with progress reports, meeting minutes and plan changes. Written documentation helps make...

...plasmid, and can be assayed conveniently. A direct test for your gene of interest may be another method ...always mission critical to make sure the plasmid replicates in the cell, as many of these experiments are... 100-400 ug/mL *Not comprehensive.   ** In eukaryotes.   ***The concentration used for selection is...

...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...

...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 𝛽-galactosidase. Complementing...this deleted portion, for example on a plasmid, creates a functional 𝛽-galactosidase. Blue-white screens...

...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...

... 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 (...

... both on and off target. Across all loci tested, error rates overall ranged from 0.3% to 10%, in comparison...Importantly, this is true in both mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the...

...be used to express proteins. Viral expression cassettes typically contain a promoter (inducible or constitutive...lineage, and so much more! Libraries allow you to test thousands of elements simultaneously in your system...dose. Vaccine development strategies. Image courtesy of Gorry et. al, 2007.   Clinical applications...

...to the right indicates a higher signal, or upregulation, and a peak to the left indicates a lower signal...continue gating populations of interest with normal gates, just based on a histogram plot instead of a dot...

... packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus using different...post‐transcriptional regulatory element; sequence that stimulates the expression of transgenes via increased nuclear...U5 and the rest of the provirus. 3’ LTR Terminates transcription started by 5' LTR by the addition...

...companies or check the literature. Pro tip! Early gates, like your live/dead markers or commonly used markers...specific cell populations, often used in the later gates, are more likely to require FMOs. Isotypes Isotype...rather than for setting up your initial voltages and gates. Ashlyn has a research background in CD8+ T cells...

...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...

...calculated (details in [2]) and can be used to quantify rates of mutation in a bulk or single cell-derived genomic...instructions on how to design such assays, in addition to testing their performance (2). Remember that although digital...transfection of a novel TAL effector nuclease (TALEN) facilitates efficient knockout of HIV co-receptor CCR5. Nucleic...

... trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its target in cis and cleaves... used by Jinek et al. 2012, or browsing for the latest AsCas12a plasmids, Addgene and CasPEDIA have got...from CasPEDIA.   How can I use it? CasPEDIA operates like many search engines, featuring both a general...

...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...

...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...

...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...

...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...

...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...

...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...