We narrowed to 1,042 results for: URE

...This blog post is the first in a series that will feature advice for students, postdocs, and young Principal...their top 3 tips. Tom Ellis 1. Keep up with the literature. He recommends setting up a few Pubmed email ...“If you don't read, you're doomed.” 2. Always be sure to include various positive and negative controls...

...paper that appeared in Nature Biotechnology back in 2011 and then in another Nature Methods paper last year... far-red light. As the researchers described in Nature Communications last year, “The capability to control...

...get it into your plasmid backbone, and zap the mixture of DNA into cells. Unfortunately, not every cell...post, but don’t worry, we’ll cover screens in a future post. Positive selection: Cells that have gained...toxin genes, and the sacB gene). By tapping into nature’s toolbox, scientists are finding more and more ...

... distinct C-bend to escape possible danger (see figure). Larvae also perform spontaneous slow swimming...distinguish between swimming behaviors, the researchers measured the tail movements of 5 day old fish. The larvae...behavioural module in the vertebrate spinal cord." Nature 461.7262 (2009): 407. PubMed PMID: 19759620. PubMed...

...and a drug is developed, it’s good practice to measure the specificity of that target and validate the... is where CRISPR can save time and money to make sure quality candidates are selected.      CRISPR...discovery pipeline.   CRISPR and drugs in the future While CRISPR has already done a lot for drug discovery...

...of soundwave pressure required to collapse them: arg2 vesicles collapse at a lower pressure than arg1 vesicles...noninvasive imaging of microorganisms in mammalian hosts. Nature, 553, 86-90. PubMed PMID: 29300010. Additional ...

...proteins, DNA, and RNA used in CRISPR.   Figure 1: Overview of the parts of CRISPR. The bacterial...in the CRISPR array that will become guides for future targeted degradation. Before integration into the...CRISPR–Cas immunity and genome editing enzymes. Nature Reviews Microbiology, 20(11), 641–656. https://...

...archaea and eukaryotes such as yeast and plants. In nature, plasmids provide one or more functional benefits...for the life scientist or bioengineer.   Figure 1: Map of a plasmid with its elements described...constructed in the lab? Due to their artificial nature, lab plasmids are commonly referred to as “vectors...

...Zimbabwe Dr. Fiona Robertson (far left) is a Senior Lecturer and a Plant Molecular Biology and Biotechnology...manage insect pests and to support sustainable agriculture. For this project the expression of isolated .... Subscribe to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees...

...in our repository and will discuss some of the features you may want to consider as you make your choice...vector that you will need. You will also have to make sure that your plasmid has been incorporated into the...promoter. Other vectors can be expressed in fly culture cells like S2 and carry the metallothionein promoter...

...advice from the successful PIs who were previously featured in "9 Tips to Achieve Success in Academia". Important...evaluated your plan? Have you read the relevant literature to understand where the field is today? Have ...the importance of keeping up-to-date with the literature of a couple of well-known journals that discuss...

...indefinitely at room temperature (thus also saving freezer space).  The procedure becomes less attractive... agar slices.  Molecular biologists used this procedure daily before the advent of techniques like PCR...

...selective media, and provides those bacteria with a pressure to keep your plasmid. Viva la (bacterial) resistance... antibiotics against Gram negative bacteria. In future posts, we'll detail selection in non-bacterial ...strains have natural antibiotic resistances, so make sure your plasmid and E. coli strain are compatible! ...

...successfully reprogram. Unfortunately scRNA-seq only captures the gene expression of cells at a single time ...analysis and the remaining cells were maintained in culture. iEP cell physiology and reprogramming efficiency... lineage and identity in direct reprogramming." Nature 564.7735 (2018): 219. PubMed PMID: 30518857. Guo...

...the public in a meaningful way, and not solely to ensure a profit. Often, these companies believe they possess...chance to carry out a comprehensive review of the literature informing the notion of “heading” in soccer as... of my thesis: a survey instrument designed to measure how individuals respond to issues of unsettled ...

...the foundation of all good communication is making sure everyone’s on the same page. So for our first post...education is education that is provided through a structured environment such as the classroom or a defined...specific audience - giving consistency, purpose, structure, and intent to the messaging - while also creating...

...colleagues and is invested in improving research infrastructure, from our educational materials to our repository...and ScFvs, based on community demand. Figure 1: Antibody against PSD-95 from the Trimmer Lab...Reproducibility: Standardize antibodies used in research. Nature 518:27–29. https://doi.org/10.1038/518027a  Resources...

...seedlings or callus (undifferentiated plant cell culture), which is great for a wide variety of applications...found in all other plants, anthocyanins. Figure 1: An adult plant expressing RUBY. Photos by Andrew...can be selected without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing...

...stressors, including emotional stress or changes in temperature.  “The disease was first described in 1971,” ..., it is still sometimes wrongly diagnosed as a seizure disorder.” In 2012, researchers identified the ...Thailand will receive much better health care in the future based on our studies.” Thank you to all the BeHeard...

...that we aren’t still pitching for feedback.  Your venture should continuously evolve and respond to feedback...number of experts, including Harvard/MIT faculty, venture capital and angel investors, and business development...idea has potential to make a difference, then make sure you’re able to answer every possible question related...

...model organism for a few reasons: Affordable to culture and easy to work with Grows extremely fast (doubling...induced by removing methanol from the media. This feature makes the strain tractable for peroxisomal studies... for genome engineering. A unique evolutionary feature of K. phaffii, relative to S. cerevisiae, is that...

...receiving only 1 TF expression construct. Cells were cultured following a differentiation protocol before performing...so that a single TF was delivered to each cell, future applications could deliver multiple TFs to each...on the Addgene Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral Transduction...

...discuss those ideas and build from those.   Figure 1: Virtual lab meeting? No problem! You can still...final screen can also be saved and exported as a picture. I believe that showing something different from...document can be shared and serve as a springboard for future discussion. Collaboration and creativity are pivotal...

...got along well, suggest a coffee chat in the near future. Even just one more meeting (in person or virtually... but I’ll never forget the strawberries. Grand gestures are memorable! Attend events for one organization...defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is actually a map of Joanne...

...addition to thorough ethical considerations, in making sure that the research has been adequately proven to ...unethical behavior (it’s hard to argue otherwise). To be sure, throughout the history of medical research, there...on necessary actions to review and decide on the future of genome editing technology. We must stop punting...

...formats. For example, several journals like eLife, Nature, and Cell use  Research Resource identifiers (RRIDs...resolvable compact identifier. Addgene works hard to ensure that materials available through our repository...Addgene Blog Read a blog post on Our Citations Feature Check out this post on Scientific Reproducibility...

...that are then barcoded for identification. This mixture of barcoded fragments is known as a *library*. ...additional benefit of plexWell™’s sequential tagging procedure, the quantities of library molecules for each ...multiplexed pool. The cost of these normalization steps, measured in time and consumables, can be daunting for large...

... the library, use next-generation sequencing to ensure that the library has retained gRNA representation...produce high titer lentivirus from the library, ensure gRNA representation in the viral prep, infect target... you're interested in studying. You need to make sure your assay is scalable and robust. What you don'...

...they also uniformly stepped back to give a big picture perspective: Dr. Yamanaka talked about his clinical...trial patients with advanced macular degeneration; Laurenz Studer discussed his work with Parkinson's Disease...presenters, it was gratifying to see that Addgene has ensured that plasmid acquisition is not a rate-limiting...

...predict protein structures, then you might know a thing or two about computer modeling. Make sure to focus on...? Or perhaps you have presented in some public lectures? These transferable skills could be meaningful...

...see the Introduction to Fluorescence Microscopy lecture at iBiology. Commonly used filter sets & relevant...this filter set and have reported brightness measurements. Here, EGFP outperforms the improved folding...variants, presumably due to the lower growth temperature. This also suggests, however, that there is no...

...virulence. Scientists have studied how HGT occurs in nature and have learned how to introduce genetic materials...Transformation, transduction, and conjugation occur in nature as forms of HGT, but transfection is unique to ...material from the environment by bacterial cells. In nature, this genetic material often comes from adjacent...

...genes for “gas vesicles,” a gas-filled protein structure that water-bound photosynthetic organisms use ... check out this blog post. Bourdeau RW, et al., Nature. 2018. PubMed PMID: 29300010. miRNA reporter vectors... date, containing all 620 sequences of probable mature human miRNAs. Weiss’s collection consists of a...

...screening blog post for more information.   Figure 1: Result of a blue-white screen. Image from Stefan...more about colony PCR in this blog post.   Figure 2: The steps of colony PCR.   Sanger sequencing...in this case a plasmid. Sequencing is one of the surest ways to know that your insert is what you expect...

...molecules of the RNA backbone to increase stability.  Nature has evolved ways to prevent degradation of important...and rendered useless after less than a minute of exposure to the appropriate light. The photoactivatable...to engage a target sequence. Upon a brief light exposure (several seconds) the gRNA is released and the...

...consolidate several steps into one. Soon you’ll figure out what prompts you need to consistently and accurately... accurately run a protocol.   Figure 1: Evolution of a Western blot protocol, from a full page ...experimental steps are worth doing, and doing well, so make sure you’re taking time to do them carefully and thoroughly...

...results you've measured should change established theory. So, what can you do to ensure that your skills...our website and social channels to apply for our future events. In addition to this annual event, graduate...

...additional flexibility in pseudotyping.   Figure 1: Genome of a prototypical gamma-retrovirus.  ...nucleus through the nuclear pore complex. This feature may be useful for the transduction of terminally...-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to be descriptive...

...and you think there are four important times to measure that growth rate, you report the results of all...between hypothesis testing and generation, can be sure that results will be reported regardless of outcome...the journal and reviewers are incentivized to make sure that any possible null is really a true null and...

...processes directly affected by light. Long-term light exposure can also lead to cellular phototoxicity, and experimental...substrate like coelenterazine is supplied in the culture medium to allow RLuc8 to produce photons. The original...wavelength to specify which FP emission you’ll capture. Since the Nano-lantern system does not use external...

...won’t work if the fragment ends have secondary structure or too much sequence similarity. REPLACR-mutagenesis...expressing recombineering proteins Red/ET from a temperature-sensitive plasmid. Homologous recombination then...also well suited for insertions and deletions. The figure above shows guidelines for primer design; a desired...

...at all 1,500 or 1,600 transcription factors and figure out where they bind to the genome. We’ve been doing...other scientists quickly? Myers: We were hoping to figure out how to do this for many hundreds of transcription...as well. We would also like to put some social pressure on many other labs that may be doing something...

... known as the resting potential (Vmem), can be measured as the imbalance of electrical charge between ...of the pHLuorin fluorescent protein. This GEVI features increased brightness and improved plasma membrane...Levin, Director of the Allen Discovery Center. The future of this field has a high potential for the advancement...

...Find the CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed PMID: 30377362. Studying ...the cell membrane where cell voltage can then be measured. Archon1 is  bright, exhibits good localization...processes in vivo. Many photoactive proteins exist in nature, but developing them into optogenetic tools can...

...These workshops illustrate the open and modular nature of the workshop content: different parts could ...for securing funding to support this effort. The future of reproducibility workshops and lessons learned.... Be on the lookout for upcoming workshops at future conferences this year. Workshops are confirmed ...

..., with the expectation for major advances in agriculture, materials, chemicals, and medicine. As they’...and where all of it is likely to lead us in the future. Building circuits from CRISPRs Addgene: What...interactions. That’s relevant if you want to make sure you don’t target anywhere else, but for synthetic...

...increased pressure of the added liquid. The volume should be minimal in order to minimize the pressure into...mouse breeding colony) Environment Olfaction, temperature, humidity, noise level, circadian effects (inactive...

...Golden (Gate) pot (Pryor et al., 2022).     Figure 1: Workflow for high complexity Golden Gate Assemblies...cycling conditions (see Table 1).   Enzyme Temperature Incubation time/cycling Overhang length ...also optimized the thermocycler protocols and made sure that T4 ligase was the correct choice for the job...

...IgG2a antibody. When using a monoclonal antibody be sure to use a highly specific secondary antibody, rather...IgG2a > IgG2b > IgG1 (Manning 2012).      Figure 1: Broadly reactive versus subclass-specific antibodies...antibodies rather than generally reactive ones. And make sure to check for any reported cross-reactivity when ...

... have reached labs in over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image... the spike protein was developed based on 100 structure-guided spike designs and has improved protein ...Maynard JA, Finkelstein IJ, McLellan JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-...