...preparation approaches described previously require a preliminary double stranding step. The double stranding step... library preparation procedures. This finding eliminates the double-stranding step and reduces both the...confirm the identity of AAV genomes and detect contaminating AAV DNA. In addition, we wrote an open-source...uses the VGS data to determine the percent of recombination present in Cre-dependent vectors and a script...

...sequences on a daily basis, many found these tools inadequate for planning, visualizing, and documenting their...elements such as promoters and transcription terminators, the boundaries are less obvious. We found that.... In an effort to be rigorous, we dug into the original literature, some of it decades old, to provide...mismatches, our algorithm could annotate a feature inaccurately. The best example is fluorescent proteins, which...

...electrical activity through imaging. Since it’s illuminated by red/orange light, Archon1 can also be used...Chimeric channels for neuronal manipulation Recombinases Plasmid Serotype Name 50363  AAV5  AAV ...labeled neurons because background fluorescence from inactive neurons is reduced. 105322  AAV1  pGP-AAV-...densely-labeled neurons because background fluorescence from inactive neurons is reduced.  Additional resources...

...disturbing the tissue for imaging or collection could contaminate it. I have used it for selecting transformation...promoter with self-cleaving 2A site linkages and a terminator. Image from He, Y., Zhang, T., Sun, H. et al....untransformed seed from the same plant beginning to germinate. (Top right, second panel) Transformed seedlings... Biology Department at the University of North Carolina at Chapel Hill. He has a passion for microscopy...

...the first vaccination and after both vaccinations. For the B.1.351 variant, a single vaccination didn’t ... SARS-CoV-2 variants coincide with increased vaccination efforts. So, how effective are the current vaccines...

...particles. A single viral particle is called a virion. Finally, viral vectors, which are used in the lab, are ...contrast to bacteria and eukaryotes, which use a binary replication process whereby one cell copies itself...engineering controls — changes made to the virus that eliminate or reduce the risk of infection — researchers ...genetically modified to either limit or completely eliminate their replicative ability. The remaining particle...

...next experiment. Nucleic acid based machines Recombinase-based State Machines Enable Order-dependent Logic...inputs are received by a cell. These so-called recombinase-based state machines could potentially be used...development. Read this post to learn more about recombinase-based state machines and how they function. Adapting...them. CRISPR Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening CombiGEM is a technology that...

.... We will continue to provide you with more fascinating, informative, and actionable content in the coming...genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions, deletions...Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and tired of having...you’ve made the right one. In this post, Addgenie Melina Fan guides us through the basics of the validation...

... embryonic stem cells, they found very high recombination percentages (78-79% after two rounds of transduction...application of iTOP Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to see if iTOP ...events. For these reasons, iTOP is best suited to binary systems in which transient protein expression has...long-lasting effect, such as Cre- or Cas9-mediated recombination. Both primary hESCs and immortalized cells treated...

...many cases, you won’t see any signs of DNA in your final tube after purification. How do you know if you ...your DNA prep and whether there are any other contaminants. Nucleic acids (DNA and RNA) absorb maximally...method also gives an indicator of DNA or RNA contamination based on the presence of other bands or streaking...methods, this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary electrophoresis...

...projects, we offer classes, workshops, book clubs, seminars, and social events. Building a community science...community projects. Recently, Timothy Stiles and Mrina Nikrad have started the Open Yeast Engineering Project... not be pursued in an academic or industry lab. Mrina Nikrad, Ph.D., is an expert in microbial ecology...! Dennis Oleksyuk, Raphael Laurenceau, and Tami Pudina have initiated a soil microbiome project. This ...

...annotate things as they come up in their minds. The final screen can also be saved and exported as a picture...miscommunication, a lack of leadership, and inappropriate tools or writing approaches can lead to frustration...frustration, delay of publication, or even the termination of a project.” Take brain breaks Meetings can...Wuchty S, Jones BF, Uzzi B (2007) The Increasing Dominance of Teams in Production of Knowledge. Science 316...

...I’ve always been fascinated by the human body and how it works. I used to stay up past my bedtime, poring...baby’s health but adamantly refused to get them vaccinated. A classmate told me that they never finish a...more likely to rate that study as untrustworthy, inaccurate, methodologically flawed, and biased. As a result...information gained from that research remains inaccessible, behind paywalls and in technical jargon that...

...nearly all cell types in the body from otherwise terminally differentiated cells simply by expressing four... a community largely created due to Yamanaka’s seminal work. It didn't take very long to realize that...Hideyuki Okano presented ground-breaking work on spinal cord injuries; Jessica Young shared research on..., a half-brother who is a quadriplegic due to a spinal cord injury, and numerous family friends who had...

...recent years, there are many fluorescent protein combinations that can be imaged together, but this also means...microscope(s) you intend to use. An accurate determination of whether two fluorescent proteins can be separated...Cy5 (e.g. Semrock's) and the similar 4-laser combination on a confocal (405 / 488 / 561 / 640 nm). In ...try several proteins in your system of interest. Finally, in this set of proteins the green and red proteins...

...backbone with a T7 promoter driving multiple combinations of WT, codon optimized, untagged, cleavage sites... sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...found in the bacterial expression plasmids. Each combination is present in two sets of backbones which drive...expression system. Mammalian expression plasmids Finally a set of wild type and codon optimized, full length...

...antimicrobial.  Conjugates And HRP brings us handily to the final topic of this post, the conjugates, or signaling... a brief introduction to the wide, wild, and fascinating field of signaling molecules. Assay Type Westerns..., simple to use, not sensitive to light in its inactive state, and cheap. Unless there’s a specific reason... on Addgene.org Find plasmids encoding recombinant antibodies at Addgene Topics: Antibodies...

..., but it’s also found in tissues. IgG is the predominant isotype during a secondary immune response, which...switching is caused by a specialized type of DNA recombination. Heavy chain constant genes are clustered together... Figure 2: Isotype switching requires DNA recombination of the antibody gene locus. B cells start by...heavy-chain constant domain gene cluster. DNA recombination can delete intervening heavy-chain constant ...

...continue to produce the specific antibody that the original B cell produced. Because of this, all of the antibodies...hybridomas producing the most specific antibodies. Finally, the selected hybridoma is expanded as a clonal...reproducible experimental results. When you are originally selecting a hybridoma, you can choose one that...from the hybridoma into a plasmid to create a recombinant antibody.     Figure 2: Changes to the...

...the page. Finally, as previously mentioned, enlist the help of your network to polish the final draft prior...don’t guess. Simply reach out to the program coordinator with your question(s). This precautionary step...notice within 24 hrs, send a note to the program coordinator.   6. Virtual interview prep Prepare for your...