...plasmids, they are generally electroporated into electrocompetent cells to ensure the highest possible transformation...use for your transformation(s). The amount of competent cells you need total and per transformation may...liquid culture, all of the bacteria can grow and compete in the same container, making it easier for a smaller...be its own colony, and will not be in direct competition with other colonies containing smaller, recombined...used for libraries that are similar in size and composition as examples. Table 1 provides an overview of ...

... pooled CRISPR library is a complex mixture. Pooled libraries are composed of many different gRNA-containing... and versatility. CRISPR/Cas9 consists of two components: the non-specific endonuclease (Cas9) and a single...any genomic locus, provided the target is unique compared to the rest of the genome and located immediately...distribution is analyzed in the drug-resistant population compared to a non-treated control group. In this scenario...

...one such technique and has been used to produce complex fluorescent reporter constructs. These fluorescent...gene-expression sequences, and more. By combining these components, one can generate constructs to visualize cell...terminus or the 3’ terminus of the ligation product. Comparison to other high throughput cloning strategies There...modules, because neighboring fragments require compatible cohesive ends Engler et al., 2008 Gibson Cloning...facilitate the construction of plasmids with many components. The creators of the MXS-chaining method have...

... takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. ...PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based kit, this ...this did not reduce cloning efficiencies.  When compared to fresh columns, regenerated columns had similar... reduce the number of spin columns you need to complete your research. Do you know of other nucleic acid...

...will link biomolecules to the gel and add gel components to polymerize. Step 3: digest the sample to allow...labels you could use. But many ExM protocols are compatible with conventional antibodies, chemical dyes, ...and least flexible one. Tissue expansion is not compatible with in vivo applications and there’s just not...isotropic expansion depends on how well the gel components penetrate the sample, how well the gel polymerizes... valuable tool, especially when paired with complementary experimental approaches. Since it arrived on...

...you’ll want to ensure that your buffer components are compatible with your conjugate and conjugation chemistry...the assay. Where possible, it can be useful to compare recombinant antibodies to their parental hybridomas...majority of antibodies are still equally effective compared to those aliquots frozen immediately post-harvest...Chen E, Haidar JN, Srivastava A, Goldstein J. Comparison of predicted extinction coefficients of monoclonal...

...feel comfortable speaking on topics that would compliment the STEM curriculum. As it happened, I’d met ...simplistic and I would insult the students. Too complicated and I would quickly lose their interest. I won...careful to keep it relatively simple. I honed the complexity of the talk by running it by the teachers and... For CRISPR, I focused on the fact that, like computers and the internet before it, genome engineering...posts Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

...collaborations, and commitments to and proof of compliance with the grantor’s requirements as well as state...reports are submitted and project evaluations are completed. To enter and advance the profession, you can ...Professional Development Strategy to increase your competencies and skills in the grants profession. Your professional... and the candidate’s knowledge of the 8 core competencies through a multiple choice exam.  The bottom ...grant professionals and to build your skills and competencies. Becoming a grant professional is rewarding ...

...into a translated protein. As depicted in the accompanying cartoon, they have a multitude of uses including...TAP) once referred specifically to a combo tag comprised of a calmodulin binding peptide (CBP), a TEV cleavage...ProtA IgG-binding domains. TAP has since come to encompass several other tag combinations, though frequently...and Gly residues   This article is not a comprehensive guide to all tags, but rather a quick overview...Biotechnology Journal 2012 , (7): 620-634. Full Text A complete list of free handbooks for protein purification...

...fields in order to provide useful solutions to complex challenges. As a cross-disciplinary researcher,...Many of the challenges in science today are too complex for any single discipline to solve them on their... disciplines leading the way, the future of complementing published research with video overviews was ...may never access online journals or even use a computer.  Currently, two of the more popular biology videos...derived from research. The platform uses video to complement traditional print journals and expand our ability...

... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...-tag and, more recently, OLLAS (Escherichia coli OmpF linker and mouse langerin) and Sun Tag (Viswanathan...in order to be expressed stably in neurons. As compared to epitopes tags, FPs are intrinsically fluorescent...fluorescent proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags. Indeed,... structure resembles images of the deity (see comparison image above). The church of the Flying Spaghetti...

...brainstorm session is to start with something completely unrelated to the issue and consider how it could...unique solutions to the same problem. If your lab/company/team is still working towards a culture of encouraging...just as valuable! In 2019, Addgene implemented a company-wide New Ideas Survey. Anyone could fill out the...of which, when implemented, impacted the entire company. It is important to note that the person who submitted... is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism When ...

... research compiled in the ELN. Without such references the record of research is incomplete, making it...commercialization office, the CIO’s office, and research computing. The two guiding principles emerging from the ...the record of research captured in the ELN is incomplete. Thus, to support reproducibility an ELN should...select and deposit individual documents or more complex datasets into the desired repository. They also...

...applying to the specific company or lab. Take some time to research the lab or company via their papers and...place to state why you’re applying to the job, company, and/or lab, and how you feel connected to their...apply for [title of the position] at [name of the company/organization]. I am a [adjective that describes... are interested in working for this lab and/or company. Remember to proofread your cover letter, and ...

...sgRNA targeting the genomic site of interest. We compared the pCas/pTargetF system with the updated pEcCas...system.   Advantages of the pEcCas/pEcgRNA system Compared with the original system, the pEcCas/pEcgRNA is...strategy for plasmid curing, providing users with a complete editing process. The MUCICAT system While the ...dehydrogenase expression that is 2.6 times higher when compared to the industrial strain. In our metabolic engineering... pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction...

...necessarily mean that you are a future employee of the company. It does, however, offer you an opportunity to ...like to work, get online and take a look at the company or institution’s career page, where you’re likely...openings. A good approach is to do this first for companies at which you’ve already made a personal connection...result in a recruiter giving you a call. Many companies enlist the assistance of recruiters for identifying...development center, if one exists. Get online! Review company and institution career pages. Make yourself visible...

...the ability to cleave the RNA target.  Table 1: Comparison on Common CRISPR Enzymes Name  Enzymatic Domains...Amplified nucleotides are combined with Cas 13a in complex with crRNA targeting the sequence of interest and...multiplexing; expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although... cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found that LwaCas13a ... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

...in the targeted brain region after stimulation compared to controls without Chr2. c-Fos is a reliable ... to proceed with your behavioral experiments. Comparison of c-fos amongst animals that receive different...Inhibition of neuronal activation is a little more complicated since basal neuronal activity in your brain region...writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow...applying optogenetic tools derived from direct comparative analysis of microbial opsins." Nature methods...

...revolutionised the way we apprehend molecular complexes and is still a very useful tool.    Like a guitar... These issues can be overcome by choosing a complementary pair of fluorophores. To maximize the FRET signal...to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian...Clover pGWF1 Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between...especially when you investigate unknown molecular complexes. But it is a parameter that you have to keep in...

... of the policy comprised of numerous objections and concerns. Perhaps least compelling of these were those...to individual researchers. Yet accurate and comprehensible organization of data is essential for efficient...numbers of figures such that many have multiple, complex panels. PLOS, itself, has not finalized its definition...collections and interpretation of data, publishing complete data, rather than just a single projection of ...