... and the things you need to include in your manuscript. What information do I need to include in my paper... to anyone who wishes to read it, without them needing a subscription to the journal. An increasing number...reviewers and the editor decide that your manuscript needs to be revised before it can be published, this can...

...transgene-free and vector-free cells Genome integration Need to sequence cells to verify excision didn’t introduce...genomic integration Multiple transfections required Need to sequence to verify there’s no genomic integration...-free cells No genomic integration Inefficient Need to verify loss of episome Minicircle Non-integrating...-free cells No genomic integration Inefficient Need to verify loss of minicircle RNA Delivery DNA-Free... difficult to purify the reprogramming proteins needed to perform these experiments. Assays for iPSCs...

...how many do I need? When designing a plasmid containing a polyhistidine tag, you’ll need to decide whether... binding without eluting the target protein and needs to be empirically determined. In addition, some ...Longer histidine tags are also recommended if you need a very pure sample. Because binding strength is ...

...2013). We found that the fluorescent proteins can, indeed, interfere with the correct function of the protein...this was problematic, because the DREADD receptors need to be located on the plasma membrane to be activated...DREADDs. In studies where the fluorescence is not needed, the use of HA and other short epitope tags (Kimple...receptors potently activated by an inert ligand. Proceedings of the National Academy of Sciences 104:5163–...

...the research antibodies in your refrigerator often need a little help to be useful. Mainly because, well...available, that never guarantees the antibody you need will be conjugated, or that it will be conjugated...conjugation, it is important to keep in mind that you will need to confirm your antibodies’ performance after the...which can interfere with conjugation reactions. If needed, you can often remove the incompatible agents by...

...stakeholders. However, while most institutions are indeed going ahead with their planned raises, some are...ahead with the raises as they see fit, following feedback from postdocs and faculty (at time of writing,...detrimental effect on the morale of postdocs, and indeed we have been contacted by a number of postdocs ...keep raising salaries and that we are submitting Freedom of Information Act (FOIA) requests for their postdoctoral...

...Addgene's resident blog editor, I could take the needed steps to continue where the guide left off - namely...conjugates. And, of course, once you have them, you’ll need to either re-develop or re-optimize your assay for...identified but a smaller or less immunogenic version is needed.  Single-chain fragment variables (scFvs) are ~...will arise in your research or what data you may need to collect. Understanding available alternatives...

...projects. Right now we don’t have to worry about succeeding at our core mission. Plasmid distribution is ...this and recognize it all the time. We love this feedback and are always adding to our “google doc of gratitude...awhile I worry a very little bit about hiring when we need to do it quickly. Hiring is hard but as a 3 year..., excellent intentions, or incredible drive to succeed.  Thanks to all the Addgenies and congratulations...

...is sometimes at odds with the space needed to be creative. “You need a lot of time to let you mind process...usually start out my days looking through our Twitter feeds and dealing with social messages and postings,” ...organizations have so many [science communication] needs. I worked for one company, but there were a ton ...

...designed to support new approaches that address the need to train our young scientists for futures both inside... symposium we will discuss where better data is needed to help shape and inform the overall structure ...graduates and postdocs – and from what fields – are needed to adequately supply these demands? Are there other...biomedical research from its systemic flaws." Proceedings of the National Academy of Sciences 111.16 (2014...

...of the article – via the journal (though you may need to be on a network with an institutional subscription...literature until you have all of the information you need. Reading primary research literature The best way...feasible for every paper you will ever read. So, you need to come up with an organization system that works...works for me. I can often remember which paper I need to open, but when I can’t it’s an easy search. Writing...

... to see how funds were allocated. Step 3: Get feedback from your peers and advisors Once a rough draft...reach out to peers and collaborators who have succeeded at grant writing to see what tips they found useful...reviewers Reviewers may take 6-12 months to give feedback on the application. Make sure to invest time and...the application process? What types of writing feedback do you find most helpful? Have a great grant writing...

...drop-down options for you to choose from, while others need to be filled with short-answer text. You can also...partial sequence cells. *Note: If you believe you need to deposit a collection that is greater than 75 ..., S. pombe (fission yeast), A. thaliana (mustard weed), O. cuniculus (rabbit), synthetic, or other. If...appear in your Addgene account under My Plasmids, we need to know your Addgene account username. Shipping ...

...your western and your results look… funky? Do you need to start a whole new blot? Never fear: membrane .... Dry nitrocellulose membranes are brittle, so proceed with caution. Unlike PVDF, they can be returned...antibodies. If you have already applied antibodies and need to dry the membrane for storage, but plan to reprobe...more strongly or that have saturated the blot may need longer incubation for effective stripping. Some ...

... and skew results measuring locomotion. Mice may need to be given some time to adapt to a testing apparatus...stress level, origin (vendor or in-house mouse breeding colony) Environment Olfaction, temperature, ...possible effects. Injection Parameters: The injection needle should be as thin as possible to minimize the damage... damage to the brain tissue. The injection speed has to be slow to allow the viral particles to diffuse...

...was reported, and we felt that, given the critical need for low-cost, field-based diagnostics, our approach...nights, not unlike most research, but the urgent need for the test did catalyze our resolve to get the...reach out to the greater research community when we needed additional expertise. Perhaps most striking is ...advance our programmable approach to other critical needs and to further develop our rapid prototyping and...

...optics have become go-to providers of materials needed for optogenetics. There are several important considerations...considerations with fabrication of the fiber optics needed for optogenetics. First, the length of the fiber...brings me to the second key point; the fiber optics need to be polished to significant smoothness to prevent...Before you get your experiment started, you will need a laser of the appropriate wavelength to stimulate...

...binds. For example, Myc1-9E10 binds EQKLISEEDL (pronounced “equi-kli-seed-le”). (3) Tag all the things! ...proteins of interest and proteins tagged with it needed to be chemically treated before the antibody could...recognized by Myc1-9E10 down to 10 amino acids (EQKLISEEDL) - larger than the substance P peptide, but still...

...precise expression levels. The BIOFAB project is seeding the beginnings of an operating system for programming...genetics and chemistry of life, then this new language needs to be free. The future of synthetic biology Addgene...development of enough sensor domains. Ultimately, we'll need a couple thousand or so sensor domains for everything...could imagine. That I think is the next priority; we need the keyboards and displays for control and visualization...

...rely on standard curves in their day-to-day work. Indeed, viral vector production facilities frequently ...do not know what concentration to expect, you may need to run the experiment a few times and optimize your...contaminants that could affect the measurements. You'll need to prepare the standard, standard dilutions, and...used to measure the response. The dynamic range needs to be wide enough to cover all possible concentrations...