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...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a  screen comes in. Let’s...process. This way, we can confirm the sequence of the entire plasmid. Please note that NGS verification is more...

...ancient Chinese game Go). In addition, there was an entire story-telling session organized by Story Collider...in neurobiology (and really in all of biology) require large data sets. Whether these data sets include...or sequences of microbial DNA they all seem to require some mathematical know-how for analysis. If you...from Addgene. While super useful, these vectors require a bit of expertise and can be difficult for individual...

...(FP) require external light activation, you can’t use fluorescence to monitor processes directly affected...luminescent imaging: A luminescent substrate is required for imaging, and it is important to verify that...technology. Multicolor imaging using Nano-lanterns requires a linear unmixing algorithm to separate the various...

...blessed to have an incredibly supportive Board of Directors at Addgene. Our Board helps keep our focus on ...best practices and encouraging us to explore new directions. I would like to thank them for their time, dedication...can afford to do and when. Addgene’s Legal Team acquired permission from relevant plasmid depositors and...proud to be adding viral vector services to our repertoire. Additional Resources on the Addgene Blog ...

... nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during cell division...gamma-retrovirus are so closely related, their required components are very similar. While plasmids containing...specific cell type doesn’t always lend itself to direct application of virus. However, the cargo capacity...infect dividing and non-dividing cells and doesn’t require genomic integration to express its cargo, in fact...

... AAV variant that efficiently infects neurons, directs high-level transgene expression, and has retrograde...any genetic tool delivered by the AAV.   AAVrg requires high titers for efficient retrograde transport...dye-based neuronal tracing reagents, AAV particles require receptor-mediated entry into the cell. This means...inject a recombinase-dependent chemogenetic vector directly into the region where the cell bodies of the projection...

...used. All of these expression control mechanisms require a specific modification of the viral vector for...of interest, for example. Some customization is required, but many viral constructs have already been built...applications. These libraries include sgRNAs targeting the entire genome, overexpression of all human transcription...delivery of the actual virus to which immunity is desired. Instead, components of the virus are now delivered...

...membrane so DNA can be introduced directly into the cell. Microinjection requires the use of a fine needle to... bacterium’s genome through homologous or site directed recombination. Transduction is a common tool used...donor bacterium to a recipient bacterium through direct contact. The donor bacterium contains a DNA sequence...

...population. The benefit of beads is that they do not require cells from your experiment, and they are non-specific...., CD4, CD8b, CD11b, CD19), are less likely to require an FMO. Markers that define more specific cell ...often used in the later gates, are more likely to require FMOs. Isotypes Isotype controls are negative controls... populations, but others do not. Some journals require isotype controls, while others don’t. However, ...

...students in real-world lab activities? I’m trying to inspire them and also keep up with all the new biotechnology...being asked in MIT’s research labs, BioBuilder inspires students and teachers to apply engineering principles...breathalyzer to pesticide breakdown, production of fire-resistant housing insulation, and cleaning up salt...BioBuilder's open-access curriculum available on Amazon Inspire.    Many thanks to our guest blogger, Mary Tamer...

... paper for publication in a scientific journal requires several considerations. First, it is important...; the reviewer’s name and affiliations, and the entire referee report are permanently attached to the .... If the reviewers suggest that the manuscript requires revisions, the authors can publish a new version...with the journal’s house style and check that all required information is provided. The clearer you write...

... “make the ask” once you have found someone you admire and want to learn from. Check out Joanne's Reddit...learn. Mentors can be senior to you or peers you admire.  If you see someone who gives a great talk, ask...path you are interested in following? Someone you admire for their insight and way of expressing it in meetings...locally. Your university probably has an alumni directory. Alumni who enter their information in these searchable...

...programmed to bind to essentially any DNA sequence desired. This technology allows you to create a pair of...knock out a gene. One can also exploit homology-directed repair by flooding the cell with a mutant template...to build a DNA-binding domain to recognize any desired 15 base pair sequence in the genome in rapid fashion...to create zinc finger nucleases that will bind desired sequences. TALENs are much more reliable. If you...

...is tested following each immunization until the desired level of antibodies is reached - typically within...within 2-4 months. After the animal has the desired level of antibodies in its bloodstream, the antibodies...recognize other antibodies) in experiments that require multiple antibodies that each detect a different...Once an animal has undergone a terminal bleed to acquire all of its serum, there is no more of that exact...

...is the output. The input, plasmid copy number, directly influences gene expression. By itself, increasing... the promoter region, which is predicted to be required to decouple gene expression and plasmid number... example, the synthesis of deoxychromoviridans requires three genes whose proteins are needed sequentially...ideally be optimized to promote generation of the desired product while avoiding the accumulation of toxic...

...contributed by guest blogger, Gary McDowell, executive director of Future of Research. On December 1st 2016, many...salaries - in some cases reversing these decisions entirely. At Future of Research, we had been collecting...are going ahead with raises anyway, including the entire faculty of the biochemistry department at the University...McDowell! Gary S. McDowell is currently the Executive Director of Future of Research. He is interested in advocating...

...In a recent law review article we received some entirely unsolicited praise: “Addgene, for one, is a sterling... 3.5 Addgenies strong. The Addgene Board of Directors is mature and poised to support our growth. Thanks...outgoing Chair) and Dave Root (incoming Chair) and our entire supportive Board, we have a robust committee process...referrals. Our long reaching reputation helps us hire great people from farther away via connections and...

...secondary antibody you should use when performing indirect immunoassays. The Fab regions, on the other hand...hand, are the “arms” of the Y and comprise the entire light chain and a variable and constant region of...to detect a Fab fragment, it either needs to be directly conjugated to a reporter or you need a light-chain...specific conjugated secondary antibody. These requirements limit the flexibility of and demand for Fab ...

...different mechanism thought to be associated with bidirectional transcription. In an uninfected cell, dsRNA ...interspersed nuclear elements can generate dsRNA by bidirectional transcription or as inverted repeats. These ...genomes which can produce dsRNA as a result of bidirectional transcription of their circularized DNA. These...dsDNA molecule, it does so at the expense of the unpaired DNA strand. A ssDNA molecule exists within the...

... some 400 molecules that have been shown to be required by one or more NRs for efficient and timely activation...Primary or core coregulators (those that interact directly with NRs) exist in steady-state complexes with...the new version of the NURSA website will feature direct links to listings of available plasmids for our...relationship with Addgene for the benefit of the entire research community. Thank you to our guest bloggers...

...engineering for each loci, CRISPR/Cas systems only require changing the programmable guide RNA. Gene activation... “first generation” CRISPR activator. While it requires a relatively simple construct, it exhibits modest...activation, dCas9-VP64 is great for experiments that requires modest gene activation. Synergistic Activation... to other notable CRISPR activators is that it requires a fusion protein, rather than relying on a two-component...

...heard from friends that Scientist positions often required a one-hour presentation about a prior research...manager or recruiter what presentation format they require. Frequently they will ask you to present about ...similar answer in a following interview. Expect to be tired once the interview is complete Industry interviews... interviewers might.  You’ll probably still be required to give a job talk for a remote interview. Fielding...

...that the artifactual PSCs commonly observed with direct stimulation are largely absent in the soCoChR-expressing...BBa_J23100) and expresses N-demethylases and other required enzymes, including a glutathione S-transferase... ΔguaB E. coli are missing an essential enzyme required for conversion of xanthine into guanine, and without...choice for FRET assays where a dark acceptor is desired due to its very low quantum efficiency. ShadowY...

...reporters specially designed for the aims and requirements of in vivo studies. A particularly useful sensor...or the axon (GCaMP6 only). The genetic elements required for Drosophila studies have been added for jGCaMP7...various AAV serotypes, researchers can precisely direct the calcium sensor to specific tissues or even ...molecular neuroscience. 2014 Nov 18;7:88. Tian L, Hires SA, Looger LL. Imaging neuronal activity with genetically...

...formation via NHEJ, precise modification with Homology Directed Repair (HDR), CRISPR activation or repression,... edits compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades CRISPR-based HDR ...To utilize HDR, a donor template containing the desired sequence flank by homology arms must be delivered...with CRISPR components. However, this sometimes requires intracellular injections into the cytoplasm or...

...be dull and painful to sit through (imagine the entire audience playing Candy Crush or reading email).... interviews. The idea is to ask questions in a directed way to bring out these interesting stories and... happens, but get things back on track if the direction of the interaction is not interesting to the audience...politely but firmly get them to stop talking and redirect the conversation. The person you cut off might...

... a derivative of Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors...part of the Class 1 CRISPR family, which means it requires multiple Cas effectors to carry out genomic editing... or just random mutations if the chromosome is repaired with all the original DNA. With Cas3, you don’...Mashimo, T. (2019). CRISPR-Cas3 induces broad and unidirectional genome editing in human cells. Nature Communications...

...(Are the inserts of different sizes?), and the desired coverage for the library (How important is it that...  The most important factor to consider is your desired library coverage, since this will help decide what...determined experimentally. For larger libraries that require hundreds of nanograms of DNA, you will likely need...bacteria will be its own colony, and will not be in direct competition with other colonies containing smaller...

...Potential PIs Even if you’re applying to a school that requires its students to do rotations in different labs...to introduce yourself to a few members of your desired department during the application process. Many...However, an online meeting with members of your desired program will allow for you to have the time and...is right for you outside of just the program requirements: by talking to current students about the work...

... steel wire into a backbone structure of a protein, based on its atomic coordinates. These wire models...This post was contributed by Tim Herman, director of the MSOE center for BioMolecular Modeling and the...our guest blogger Tim Herman! TIm Herman is the director of the MSOE center for BioMolecular Modeling and...