...for example) on each colony in order to screen for positive ones.  -- OR -- If your HR template has ...4 of the Cong, et. al, 2013 Science paper. For example, if you want to use double nickase, you could express...selection marker to select transfected cells (for example, plasmid with a puromycin resistance cassette, ...system with your homologous recombination (HR) template, you could then: Confirm your HR by doing Restriction...

... sequencing and RNA sequencing of complex heterogeneous RNA samples. Furthermore, such high processivity...fields of biology and enable the conversion of RNA templates to DNA but suffer from several important shortcomings...probability of the polymerase continuing to copy the template rather than dissociating).  MarathonRT, isolated...structured HCV genome (~9.6 kb of highly structured template!). MarathonRT also faithfully transcribes RNA ...

...method kills the transgene free individual. For example, with CRISPR-Cas9 gene editing, transformation ...GUS staining or are not suited to studying large samples. RUBY Red But recently, something new has been ... events and am currently planning to use it to simplify my work with an Arabidopsis mutant that can’t ...unfamiliar with these mutant plants. Using RUBY to simplify the identification of these mutants will speed...Other innovative uses of RUBY are possible, for example as positive selection of transformed plants to ...

...until you don't, of trying to understand your completely unexpected findings, or understanding why your...thrice-failed experiment, this paper was actually from a completely unexpected finding in a different experiment,...your conflict, you’ll need two other things to complete your story: an introduction, and a resolution.... big-picture problem, by asking your audience a simple question, like “Have you ever thought about how...conflict to get your audience invested in the more complex story, and then build on that in your resolution...

... learning behavioral optogenetics in order to implement the technique in my lab at The Rockefeller University...physiology in order to directly show that, for example, the firing of a specific type of neuron in a specific...too but that’s what collaborators are for). For example, if you study neural pathways involved in addiction... proper controls are required and include, for example, confirmation of opsin expression in the intended...region you wish to stimulate 2) Fabrication and implantation of the optical fiber into the region of interest...you may need to allow the virus to express. For example, if you inject into the ventral tegmental area ...

...cellular level. The reason for this is cost and complexity – anyone who works in the lab knows how expensive...expensive tiny amounts of reagents are, how complicated (and again, expensive) the different machines are...students to use in the classroom because the students simply need to add water to express a variety of proteins... engaging for students. In our BioBits kit for example, we’ve designed FD-CF experiments that result in...Addgene.org Browse our protocol videos to learn simple lab techniques Check out our molecular biology ...

... bulk of which you can find below. CRISPR for complex genetic diseases As CRISPR technology becomes increasingly...using it to investigate potential therapeutics for complex genetic diseases. So we’re revisiting this post...look at CRISPR’s potential for use in treating a complex genetic diseases, Alzhiemer’s disease (AD).  Troy...diseases with real phenotypic effect.” “It’s a very simple technology,” Doudna further explained. “That’s ...Gene Therapy: Applications, Limitations, and Implications for the Future.” Front. Oncol. https://doi.org...

...to target cells as a pooled library, which will amplify each TF to determine how its presence influences...processes. For help with library transformation and amplification, refer to the Zhang lab’s previously published...the specific dysregulation in the disease. For example, very few diseases manifest as a single mutation...expression dysregulation event. Diseases are often complex, with multiple layers of dysregulation from either...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome Library...

...was easy to find people who were interested. For example, our journal club discussion on Kevin Esvelt’s ... safety training prior to getting started. For example, Timothy Stiles joined BosLab last year with a ... space to perform early-stage experiments. For example, Dave Grawoig is using nanopore sequencing at BosLab... getting the community to collect various soil samples, extract DNA, sequence it, and analyze the data... characterize the microbiomes of the different samples. The goal is to not only learn something about ...

...equivalent of crossing off days on the calendar after completing your writing goal, which can serve as positive...graph of which days of the week you most often completed your habit is useful for figuring out when to ...makes suggestions on how to make your writing less complex. While I don’t think you should accept all of the...jargon in your writing. While it’s not possible to completely avoid jargon, this tool can help you avoid overusing...tools for grad school Hello PhD Podcast episode 15 "Simple tricks for time management: the pomodoro technique...

...make socially distanced work more creative. An example could be making lab meetings more interactive and...and brainstorm! Image from Joseph Mucira.   Implement new tools for creative interaction During Zoom...disappears from the video, and prepares a coffee, or simply moves away from the desk to switch off a few minutes...GA, O’Reilly CM, Lofton ME, Catalán N (2018) Ten simple rules for collaboratively writing a multi-authored...journal.pcbi.1006508 Vicens Q, Bourne PE (2007) Ten Simple Rules for a Successful Collaboration. PLoS Comput...

...require authors to use specific PID formats. For example, several journals like eLife, Nature, and Cell ...as Identifiers.org and N2T.net. To help authors comply with journal policies, Addgene now provides an ...these PID formats are new to you, we’ll go through examples to highlight the difference between them using...identifier and RRID formats. Let’s go back to our example plasmid, lenti-Cas9-VQR-Blast. If you scroll down...multiple PIDs separated by semi-colons. For our example plasmid it will read “lenti-Cas9-VQR-Blast was ...

...accelerating research into this virus. One such example is cell lines. The American Type Culture Collection...global endeavor. Case study 2: Plasmids Another example of reagents that have broad applications but have...dozens of times in 2 weeks. This is a striking example of the contributions that scientists are able to...popularity of the hACE2 transgenic mouse strain, for example, is sustained by the availability of this strain...damage profiling of SARS-CoV-2 and SARS-CoV with implications for clinical manifestations, transmissibility...

...take-out) or volunteering to foster a pet, for example. If you are looking for some resources specific...activities that kids can do more independently. For example, they could do something like puzzling or crafts... time.  Find learning activities for kids. For example, Skype a Scientist is now open to families at home... future experiments and other researchers by implementing reproducibility best practices in your research...helpful to you no matter what you’re trying to accomplish during this period. ...

...the Gal4/UAS system. This incredibly useful, yet simple system allows you to systematically study gene ...downstream of a tissue-specific promoter sounds simpler, this modular system saves you from having to create... making a fly line in the next blog post). For example, the hedgehog gene can be overexpressed in several...under the control of UAS enhancer sequences. For example, if you discovered five new genes and wanted to...control purposes that there are dosage effects. For example, overexpression of two genes by one Gal4 driver...

...Table 1 provides an example calculation of the weights. Table 1: Journal Weighting Example Article Number...However, determining this exactly is difficult. For example, it is very plausible that science published in...research our adjustment controls for them.   So, for example, in 2010, 10 percent of the papers with Addgene...factors that could still be controlled for. As an example, plasmid-related areas of science could be growing...

...particular reference sequence if necessary. The example plasmid on the right has a total size of 7.3kb,.... Avoid star activity. Some endonucleases (for example BamHI) are capable of cleaving sequences which ... The glycerol in the buffer will make sure your sample settles in the gel well and the dyes provide a ...you to interpret the bands that you get in your sample lanes. Choose your ladder based on the expected...approximate the DNA mass of comparably intense samples of similar size. Run the gel at 80-150V until you...

...for plasmid replication in mammalian cells, for example, and a toxin that kills bacteria may not have any...Selection methods, described below, are typically employed when creating stable cell lines and are not used...plasmid selection. Instead, reporter genes are often employed to easily monitor transfection efficiencies and...antigen (293E or 293T cells), allow for episomal amplification of plasmids containing the viral EBV or SV40...HeLa, NIH3T3, CHO, COS-1, 293HEK, Jurkat T cells Complexes with DNA; causes strand scissions 100-400 ug/mL...

...lab or pivot to a managerial role. For those contemplating the leap into management, the journey can be...redefine the value I could bring to potential employers. Over time, I discovered that I not only enjoyed...motivations, strengths, and learning styles. For example, during my time as a lab scientist, I worked with...foster a cohesive, productive environment. For example, in my current role managing a cross-functional...facilitated a team meeting to highlight their complementary strengths. By encouraging alignment on goals...

...to physically set up in the lab, RNAi is often completely ineffective or results in low penetrant phenotypes...low-cost, cell-free detection assay that can be completed without special equipment. What’s more, detection... to be easily concentrated. Viral RNA is then amplified isothermally, and detected through a toehold switch...encodes the LacZ𝛼 protein. Once expressed, LacZ𝛼 complexes with LacZ𝜔 and forms an active LacZ tetramer....modifying the sequence of the toehold switch. For example, Green has also developed toehold switches to detect...