...to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional enzyme...when expressed, is lethal to the cell. Cloning fragments are inserted into an MCS in the center of this... susceptible to or dependent on certain media components, and are rescued by genes supplied on the transformed...Expression Systems Find Empty Backbones for Your Experiments    ...

...these people you meet will review your papers or grants in the future, and don’t be surprised to know that...personally Scientists in your field will know about grants and projects that might help your work Tenure can... people. At large conferences, look for smaller events or meet-ups to connect with others more easily....conference Make sure you have a business card – even students and postdocs. There are many inexpensive/free ...

...reviewer’s comments without explaining why. If it looks like you have ignored their comments, they will...plasmids (like your Addgene plasmid #), and other reagents. F1000Research, among many other journals, has...researchers have enough information to replicate your experiments, making research more reproducible and your results...

...Scientists around the world have been making major improvements to CRISPR technology since its initial applications..., heterodimerizing domain to each of the Cas9 fragments, the authors created a photoactive Cas9 tool, ...photoactivatable repressor (dCas9). The activity of all variants was reversible and repeatable. Since these initial...in turn transcribes N-Cas9. The N- and C-Cas9 fragments then dimerize to form a functional protein due...Engineering of temperature- and light-switchable Cas9 variants. Nucleic Acids Research 44: . https://doi.org/...

...microbiome to deliver a genetic payload to the recipients using the IncP⍺-family-RP4 conjugation system...they only identified seven genera in in vitro experiments, possibly because in vitro conditions did not...The vector retains the oriT for delivery into recipients by conjugation. The labs screened for the host-range...without fixation or preparation, and deliver CRISPR components for gene disruption or CRISPRi. While designed...

...and planned out your key experiments. Unsurprisingly, many of these experiments require antibodies. You...antibody may be validated for IHC, not all IHC experiments are the same. Differences in sample processing...antibody could meet your needs.    Multiplex experiments Finally, consider whether you will be using other...

...approach or indirect approach, depending on the requirements of the specific assay. For the latter three,...indicated when the protein of interest is found in low amounts or if the conjugated signaling molecule has a weak...interest is often one of the most challenging components of any immunoimaging assay. Secondary antibodies...primary antibodies. The introduction of antibody fragments, such as nanobodies, has introduced new options...

...really appreciate the work you do to make sharing reagents and protocols so easy!” — Nina Jain, Harvard Medical...process by aligning institutions with standardized agreements and electronic signatures to dramatically speed...nominated for it, it just reflects how useful the reagents we created are for the community. More than 100...were useful for researchers wanting high quality reagents, and we realized we could use the same principles...

...of dsRNA Transposable elements such as long/short interspersed nuclear elements can generate dsRNA by ...bidirectional transcription or as inverted repeats. These elements are almost always epigenetically silenced, but... are important considerations when using these reagents. RNA:DNA hybrids Persistent RNA:DNA hybrids that...

...growth instructions (temperature, strain, media supplements), suggested sequencing primers, or mammalian ... Each sequencing service will have different requirements for volume and concentration, and not all providers..., you can use DNA-editing software to perform alignments of multiple sequences to visually check for homology...Tell us your favorite sequencing tricks in the comments section below.  Additional Resources on the Addgene...

...datasets and reagents in the field. Additional information can be found in the Datasets and Reagents sections... third phase of funding, the NURSA Consortium represents a unique funding model in nuclear receptor research...and acquiring them in a timely manner. Addgene represents an excellent solution to the problem of sharing...

..., or universities may have someone senior from accounts or HR on the judging panel. It’s important for...Skills 5. Practice makes perfect, apply for small grants too There are actually quite a lot of different... researchers, so look into applying for smaller grants from within your institute or a private funding...the experience I gained in writing small travel grants to attend conferences and a seed funding grant ...

...companies and organizations selling biological reagents have a ton of data on the catalog page. It’s not.... Although many may not realize it, all of the reagents available at Addgene come from researchers, and...don’t have the resources to recapitulate the experiments performed by our thousands of depositors. That...Google Scholar, which searches scientific articles, patents, and even some doctoral dissertations. Streamline...

...clusters without accumulation in subcellular compartments such as nuclear speckles. Furthermore, CRY2clust... distinguish real and artifactual postsynaptic currents (PSCs) this work shows that the artifactual PSCs...TagRFP-T fluorescence intensity at different time points, the authors tracked protein activity while also... assays. ShadowG is darker than other dark YFP mutants, it reduces the risk of spectral contamination ...

...labeling of neurons for tracing experiments.  Neuronal tracing experiments: While synthetic promoters with...the retina, it was more likely to have borrowed elements from a natural promoter that was active in the...labeling would be useful for neuronal tracing experiments where lower levels of labeling makes it easier...

... survive in competitive environments — like by living where no one else wants to. Some organisms manage...their host organisms to thrive in competitive environments. The genes encoding these functions form the...cryptic plasmid is among the most numerous genetic elements in the human gut. Cell, 187(5), 1206-1222.e16....

...microbiology meeting. Create an outline: Write key bullet points for what you would like to say to help build your...result, and the fundamental takeaway. Writing these points will help you remember what to say and provide ...outline you created for yourself. Hit the key bullet points and elaborate if your audience seems captivated... the project as a whole. In addition to time constraints, there may be particular poster you are interested...

...to develop therapies that could directly impact patients. And, to be honest, I’ve never enjoyed grant-writing... process can be a bit of a mystery to graduate students in academia. Getting a drug to the market usually...pages. In academia, however, writing papers and grants is an inescapable and essential part of being able...’re often required to at least attempt to write grants. If you don’t enjoy or excel at writing, an industry...

...number of cycles.  Choosing a method  As with all experiments, a number of factors will determine which approach...Resources needed Most molecular biology labs and departments will have qPCR machines available for use. ddPCR...Chung WK. Quantitative analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc... The copy-number of plasmids and other genetic elements can be determined by SYBR-Green-based quantitative...

... it can amplify DNA using only a short list of reagents and several heating and cooling steps. PCR relies...strand of DNA to add nucleotides to.  Using these reagents and a series of heating (denaturing) and cooling...cycle. However, it is common for dNTPs and other reagents to run low during the final cycles which would...of different molecular biology techniques and experiments. End point PCR’s goal is to amplify a specific...