...usurped old methods or complemented them in ways that make them much easier. A special class of FPs, the FP...functionality with a single gene Although it is possible to make a timer system composed of two separate FPs (one... This one protein method simplifies cloning and makes your experimental setup less cumbersome. 3. Portability...

...Knockout/Knock-in plasmids Gene targeting methods also make it possible to insert, or knock-in, any gene, tag...instead want to create a conditional knockout. To make a conditional knockout, researchers often use the...floxed alleles. Although CRISPR is very good at making knockouts, knocking in large sections of DNA can...

...Addgene is accelerating this research by helping to make new cancer resources easily accessible to the research...found across all TCGA tumor samples is represented, making the results of any such study as clinically relevant...Addgene's cancer collection pages Scientists are making progress. New resources and technologies become...

...about using non-homologous end-joining (NHEJ) to make deletions or homology-directed repair (HDR) to create...very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors. Addgene...downregulated, MMEJ represents another method for making targeted modifications. A recent publication by...

... does making transgenic organisms with Cas9 risk creating a gene drive? The simplest way to make a gene...Institute and Harvard Medical School. Scientists making transgenic organisms with Cas9 should be aware ...

...culture experience. Let’s go over the basics of making rAbs. Production Recombinant antibody production... because it secretes fewer of its own proteins, making the purification process simpler. Due to its high...Maity et al., 2015). If your antibody is not IgG, make sure to look up the setting for your isotype.  Antibody...

...researchers need to reflect upon before they attempt to make essentialist connections between biology and ancestral...Map (DepMap), a cancer dependency database used to make this information accessible to the research community... at all. This unexpected discovery, which didn’t make sense in any kind of biological framework, suggested...

...below). Addgene scientists inspect each sample and make sure they have been barcoded with a unique ID number...therefore particular sample is missing so that we can make a new tube from a replicate sample. The transformation...replicate and retain the plasmid. This allows us to make unlimited copies of each plasmid so that we never...

...You will often hear the term ‘grant writer’, but make no mistake, researching, winning, and effectively...writing. In fact, many grant applications and grant makers have moved away from long written proposal narratives...relationships with foundations and government grantmakers. We keep up-to-date on the changing guidelines...

... bench. Specifically, roles in technical support make excellent use of the skills developed by life science...advantageous to speak multiple languages - that makes communication with customers from around the world...convenient alternative for asking a quick question or making initial contact, but is less suitable for troubleshooting...

...zebrafish embryo develops outside of the mother, making them ideal for studying vertebrate embryogenesis...zebrafish To highlight some of the features that make zebrafish useful, I’ll outline two recent lineage...unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow” fish lines have been...

...and many of my scientist colleagues have as well. Making the transition from the bench can be jarring for...aren’t a lot of resources out there for scientists making this transition. In this installment of the Addgene...come up with a list of things to consider as you make this transition. Of course, a “desk job” could entail...

...organizers in their town hall.  Iterative improvement - Making Binning Singletons better Our experience running...also helped us find areas for improvement. We will make a greater effort in the future to match people within...Binning Singletons is its own organization, working to make scientific meetings more productive for attendees...

...amazing 111 countries (Figure 1)! But while we were making the materials available to many countries, import... to increase accessibility to our repository by making it easier to find materials and learn about them... know we will continue to ask “What can we do to make our materials both available and accessible to scientists...

...truncated forms that limit its functionality. To make biophysical and structural studies more feasible...simplicity and efficiency of this dual-plasmid approach makes it a valuable resource for advancing our understanding...assembly of the split luciferase is reversible, making PILATeS suitable for live imaging with a time resolution...

...well-researched field, you may need to get specific to make sure your article doesn’t turn into a textbook. ... short bursts (even twenty minutes was enough to make some progress). For most researchers, it’s not always... into something cohesive-- change the sentences, make it flow, and start telling the story. Like any editing...

...specific target, and the molecular processes that make aptamer biology a powerful tool. In this post, we...dependency on magnesium and higher thermal stability, making it a potentially more useful tool for in vivo studies...interaction between Corn and its small molecule makes Corn more photostable than Spinach and Broccoli,...

...Being smaller means scFvs are easier and cheaper to make because they can be expressed in bacteria, while...with other protein domains, much like lego pieces, makes them better suited than antibodies to be used as...T cell receptors and are expressed on T cells to make CAR T cells. The extracellular scFv portion of the...

...sometimes it can be a big challenge. But ask any filmmaker or videographer and they’ll tell you, a lot of...Design-Build-Test-Learn cycle adapted for video making.   Cycle One: The Ironing Board Control Center...: There were still a few adjustments I wanted to make to ensure smooth filming in the future, but we were...

...target cells with Cas9. Hooray! Now it’s time to make sure your genome edits went according to plan. In...sequence was inserted or deleted accordingly. Pro tip: make sure your PCR amplicon isn’t too large so that you...issue, you may need to select an alternative gRNA. Make sure to check out our validated gRNAs to avoid rolling...

...false positives observed in the cloning process, making it easier to produce the constructs you need. Expression...affecting its translation initiation and instead makes initiation heavily dependent upon the core of a ...to simplify the protein purification process by making it easy for a researcher to quickly generate N-...

...commitment. Another advantage is that this process makes it easy to generate wild type and mutant constructs...content, contains a lot of repeats--any of which could make the PCR step difficult--OR there is no convenient...rather than using a restriction enzyme or PCR to make linearized vector, the Cas9 enzyme along with a ...

...Becker muscular dystrophy (BMD). Dystrophin’s size makes it difficult to deliver via gene therapy, so researchers...normal function. Even low-frequency editing could make a huge difference in DMD, and it’s estimated that...Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. In vivo genome...

...2008). It can also cross the blood-brain barrier, making it useful for applications targeting the CNS (Issa...expression (Tervo et al., 2016). This functionality makes it useful for mapping neural connections. Although...depends on your experimental system. As we cannot make universal serotype recommendations, the following...

...a given phage will recognize on a host cell. To make matters even more interesting, much of what a phage...display little to no homology to anything else, making it difficult to quickly identify possible gene ...the T7 polymerase and a few proteases knocked out making them particularly useful for protein production...

...they are relatively small and easy to work with, making them the go-to choice for studying mammalian biology.... For editing the genome, the CRISPR/Cas system makes use of 3 components, a guide RNA (gRNA) of about...content in the mouse genome and the Cas9 enzyme will make a cut at the target site. This is when it gets exciting...

...characterized by large DNA recognition sites, which makes them highly specific.  Jurassic Park by Michael ...   Star Trek: Into Darkness (2013) which is the remake of the space-melodrama Star Trek II: The Wrath ...has always served as a vision of the future and to make us reframe our perspective of the current. Although...

...ve been planning, it is a good idea to pause and make sure that your antibody has been appropriately validated...guarantee recognition of the target antigen. You want to make sure that your antibody binds to what you expect... epitope information for some antibodies, which makes it hard to ensure that you have unique antibodies...

...with DIO/FLEx vectors and off-target expression making it easier to determine the spread pattern around...your cells of interest. You should now be able to make smaller adjustments to a set of injections into ...K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller I, Mitsopoulos A, Fan M (2020...

...our lab so we opted for AAV. While these benefits make AAVs useful for a wide variety of experiments, care...can still reduce AAV titer. For long term storage, make > 25 μl aliquots in 0.5 ml tubes, snap freeze in...humans and additional improvements are being made to make them even more useful for therapeutic purposes. ...