...induce recombination at pseudo- or cryptic loxP sites (estimated to occur at a frequency of 1.2 per megabase...used successfully in Drosophila, so Rubin sought to test the suitability of four additional yeast recombinases...recombination site for each enzyme, were generated to test recombination efficiency. When expressed in neuronal...each recombinase and reporter construct pair were tested for excision efficiency. B2 and R showed partial...doi:10.1007/s004270100167. PubMed. Cryptic loxP sites in mammalian genomes: genome-wide distribution and...

...mismatched recognition sites, its presence induces first opsin inversion, and then lox sites excision, therefore...Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger neural...neural excitation, whereas yellow light activates halorhodopsins to silence neuronal activity (Figure 1) ...induce DNA recombination at defined recognition sites (i.e., loxP and FRT, respectively) (Abremski and...floxed”) by two sets of incompatible recognition sites (e.g., loxP and lox511) in opposite orientations...

...from the NIH, in the United States) Strong letters of reference that attest to your talent and expertise...fellowship lasts five years. Most universities and institutes annually advertise for Group Leader positions... a postdoctoral mentor. A good mentor typically writes the grants that fund your research, guides the ...Here is my personal list:   Exercise - nothing dissipates stress and improves your perspective like intense... Jennifer Lippincott-Schwartz at the National Institutes of Health, was an Associate Professor at Albert...

...Harrington et al tested GeoCas9’s love of heat in two ways. The first set of experiments tested the ability...following heat treatment. The second set of experiments tested the ability of GeoCas9 RNPs to cleave substrate...genomic edits in mammalian cells. Harrington et al tested GeoCas9’s editing skills by transfecting HEK293...thermostability also had a longer lifetime in human blood. To test if this was also true for GeoCas9, Harrington et...

...also write test question banks, case studies, and experiments for labs, in addition to testing those experiments... at eScience Labs, an educational company that creates hands-on science lab kits and digital curricula.... [We ask them,] 'Can you read this lab. Can you test this experiment? Can you follow and understand the...working with students and faculty, Caitlin also collaborates with the sales and operations departments at...

...recombination sites. To introduce a gene of interest into the destination plasmid, recombination sites must first... fragment within the gene or loss of the gene alleviates its toxic effect. Negative selection is sometimes...and integration of the gene of interest at these sites in the plasmid excises the ccdB gene and the cells...gene is used for negative selection on 5% sucrose plates. Exchange of the sacB gene for the gene of interest...

...drop off the box at any location where the United States Postal Service picks up mail. NEB actually started...drop off the box at any location where the United States Postal Service picks up mail. Recycle plastic...it has to be packaging (no coated tissue culture plates, for example). Check the link for more details ...back. Be that person! If you use serological pipettes from Corning®, Falcon®, or Axygen®, set up a bin...in sustainability. Sign up for blog posts and updates from Labconscious. If you’re inspired by this...

...and Rep40) Rep proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins...key role in AAV genome replication. Replication initiates from the AAV genome ITRs on the transfer plasmid...in the ITRs, uses its endonuclease activity to generates a ssDNA substrate which is then nicked by its ...Alternative splicing and differing translational start sites generate the three VP proteins. The three VP proteins...alternative open reading frame (ORF). While AAP promotes virion assembly, the role of MAAP has not been...

...talks by prominent scientists, including Nobel Laureates, who speak about their work and current, trending...society currently has 1,283 members, who comprise undergraduates, graduate students, postdocs, faculty members...The society encourages interactions between undergraduates and PhD students so that they can help support...students to attend. It also hosts high level academic debates for its members to become involved with. The society...over 170 members in its first year. The society facilitates opportunities to learn about Synthetic Biology...

...year, scientists hijacked survival tactics of prokaryotes to deliver the technological biological blockbuster...demands of the biotech field, let's look at how prokaryotes originally utilized CRISPR/Cas as a powerful ...experiments exposed S. thermophilus to predatory phages to test if the exogenous DNA would be incorporated into ...these studies led scientists to hypothesize that prokaryotes developed an adaptive immune system that utilized...an interesting "arms race" between phages and prokaryotes; the ultimate victor will be the species who ...

... preparations undergo rigorous quality control testing at Addgene meaning they come ready made to accelerate...interested in faster by producing, titering, and testing viral preps for them. At the same time, we’re developing...base for the processes of viral production and testing so that researchers can use our educational resources...researcher Connie Cepko has been helping us with AAV testing and recently had this to say about the service:...virus protocols, information on how we produce and test virus, and quality control data for each of the ...

... one that represses transcription to one that activates transcription.However, this system lacks spatio-temporal...led to functional BLADE variants, too. The team tested different linkers between VVD and the DNA-binding...light, the VVD domains of the BLADE dimerize and activates transcription. The team created two constructs... cannot do with AraC,” says Di Ventura. BLADE activates transcription with the flick of a light switch...difficult to regulate quickly. Arabinose also circulates within growth media and can’t be contained to...

...validated simultaneously in the brain and the intestines of mice. The result? A very cool tool to advance...fiber implants. Due to the soft nature of the intestines and the requirement for digestive motility to...Using a programmable microcontroller, NeuroStack coordinates optical stimulation of its associated fibers ...wondering “did it actually work”? The authors first tested that the system could induce stimulation in target...excited a class of enteroendocrine cells within the intestines known as neuropods. They observed increased vagus...

... of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that it can ...following figure, in which we add EcoRI and NotI sites to Your Gene of Interest (YGOI) for ligation into... (usually 18-21bp) When selecting restriction sites, you should use a DNA analysis tool, such as Addgene...Analyzer, to allow you to identify which restriction sites are present in a given sequence. You want to choose...sanger sequencing. DNA replication by PCR has error rates that range from roughly 1 per 500bp to roughly 1...

...Image courtesy of Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested iTOP ...functions through macropinocytosis. D’Astolfo et al. tested iTOP in a series of primary cell types using Cre...Cas9 is all the rage, they had another reason to test this system. It’s difficult to determine how much....2015.03.028. PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery...

...that is, let's look at Figure 2. The blue laser excites at a wavelength of 488 nm (Table 1), which is near...line, Figure 2). The yellow/green laser, however, excites at 561 nm, which is close to the global maximum...more photons are absorbed and then emitted, which creates a relatively stronger — or brighter — signal.   ...the same laser type. Knowing that the red laser excites at 640 nm (Table 1), while comparing the two fluorophore's...is about 750 nm. If we were to use a laser that excites at 750 nm, APC-Cy7 would show a much brighter signal...absorbed and emitted by matter, and how this process creates visible colors. Next, you learned about how fluorophores...

...of pure product in an affordable way, researchers test many different combinations of promoters, terminators...Protein expression in yeast MoClo is also for eukaryotes! The technology has been used to optimize protein... chromosomal integration of larger expression cassettes (Shaw et al., 2023). Those looking to express ...cutting. Beyond sgRNAs, MoClo can also be used to test a variety of Cas enzymes with different PAM requirements...applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR kits too. The AspFlex Kit ...delivery. While Fragmid developers were specifically testing CRISPR tools, it should be easy enough for researchers...

...on a protein, enabling the labeling of protein substrates/peptides and antibodies. Example applications...intensity at ~85% of its original value after 60 minutes of illumination, highlighting its reliability for...G-protein-coupled receptors (GPCRs). Mutations in receptor sites can affect the fluorescence emission of labeled ...assay results (Hayashi-Takanaka et al., 2014)​​. Conjugates with lower DOL tend to have weaker fluorescence...., 2016). When selecting dyes, it's important to test them under experimental conditions. Common issues...toggle’ between fluorescent and non-fluorescent states in response to a specific stimuli, which leaves...

...Producing yeast expression cassettes Traditionally, generating expression cassettes for yeast transformation...in the amplification. This tool allows instant updates as fragments are added, deleted, or reordered and...Cloning sgRNAs Production of Yeast Expression Cassettes Generating constructs with multiple point mutations...typical two oligos, is more cost effective and eliminates the annealing step usually used to create a double-stranded...

...sequence and ultimately its degradation. While other eukaryotes don’t naturally have this response, they do contain...protospacer adjacent motif) in order to bind the target sites for editing. PAM sequences can be from 2-6 base ...which mediated a C to T conversion in the target sites adjacent to an NG PAM in human cells. The increased...and its variants can be used to engineer specific sites that were previously difficult. Find SpCas9 and...the Tara Oceans Project datasets. This study demonstrates the power of combining the analysis of publicly...