We narrowed to 855 results for: NTS

...same output: a spreadsheet of read counts. You can use these counts to confirm the representation of your...when you need to amplify a library for your own experiments. Library amplification 101 Library amplification... lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger. And ...is similar for most libraries. Here are the key points you’ll need to remember when undertaking library...enough DNA and competent bacteria to ensure all components of the library are represented. Typically, Addgene...need to count how many times each of the library components is found in the read list. If you also sequenced... your library. When plotted as a histogram, the counts will give you a bell curve of the abundance of ...

...undervalued.” Connie Cepko points out the limited choices in hiring graduate students and postdocs when starting...be half of my day sometimes.” He points out that the copious amounts of email are probably because a PI... than just experiments and grant writing. You have to enjoy the mentoring role.” She points out that new... having projects ambitious enough that everyone wants to recruit rather than repel and hoard.” If you'...it is once it gets fully up to speed.” He also comments that it is also surprising “ how easily people...last post in this series “Fun in the lab!”   Acknowledgments Thank you to Dr. Tom Ellis (Imperial College...

...the most recent advances and how some of these variants may be appropriate for your specific research ...nuclease When choosing the Cas9 nuclease for your experiments, there are three important factors to consider...isolated from different organisms and engineered Cas9 variants that were able to target different PAM sites (... be targeted (Hu et al., 2018).  These altered variants can allow for targeting of your genomic loci of...sensitivity in choosing the Cas9 to use in your experiments   The CRISPR drawer. So many options! ...off-site activity of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., ...suitable for in vivo research and potentially treatments. Cas12b, characterized by the Zhang (Strecker...

...other RNA components are required? CasPEDIA provides information on crRNA and tracrRNA requirements, along...field has exploded with dozens of Cas enzymes and variants available. Without a comprehensive resource, it... and cleaves ssDNA in trans. In sum, CasPEDIA documents the target substrate (DNA/RNA and strandedness...(nick, bind, cleave, mixed). This information represents the ‘big picture’ features of the enzymes documented...a candidate Cas for your experiment. Target requirements As we all know, a Cas guided by a gRNA is useless...) in some cases. The resource displays these requirements in regard to their sequence (NGG, etc.) as well...experiment. Perhaps you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly...

... interested in using GCE, here are the basic requirements of use. First, and most obviously, you need ...incorporated ncAA, ready for you do to any number of experiments with. The catch The rapid expansion of GCE led...That many GCE technologies have idiosyncrasies presents a significant barrier for those wishing to use...GCE aren’t able to successfully complete their experiments and many others are dubious enough that they ...optimization parameters the same for all three components, meaning that the final protocol may involve ...’s not uncommon to find protocols that require reagents which aren’t broadly or easily available, so you...protocol that you’ll have to rework to include reagents accessible to your lab. The solution GCE provides...

...domain from PE6e-g produced additive improvements. These variants are worth testing if you want to make...Over 75,000 pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database... potential to correct only a minority of those variants in most cell types. But prime editing, a CRISPR...goal of the experiment can all impose different constraints and affect how you select and design your prime...research, the Liu lab created and evaluated 19 PE1 variants with RT mutations known to increase activity, ...system as complex as prime editing, every efficiency counts! The optimized enzyme can be used with any of the...PE2max, PE3max, etc. PE6: Specialized prime editing variants Four years after their initial publication, the...

... of the party goers, the piñata bursts and its contents spill to the ground. There is something akin to... trouble is, with so much choice, in those few moments after the piñata burst, value was subjectively ...disconnect arising between those reviewing the contents of the piñata and the audience blindly receiving...proceed in the future. When we think of designing experiments using CRISPR, there are two major areas where... The foremost handles the design of sgRNA and represents the lion’s share of the currently available tools...example is CRISPR-GA. These tools assess repair events by type (NHEJ/HDR) and track indels at a single...more common. User interaction: Software design elements that fall into this category describe how the ...

...bind to the operator. The bound repressor then prevents transcription from occurring, which means that...), TetR preferentially binds Tet over the TetO elements, allowing transcription to proceed (pJKR-H-tetR... virus. tTA binds to promoters containing TetO elements (often linked in groups of seven as a Tet Response...binds tTA, resulting in a confirmation change that prevents binding to the promoter and turning transcription... Bujard, both the promoter and transactivator components of the system have been engineered and improved...Tet-Off vectors (Tet-Advanced)” are the most recent variants of the Tet-Off system.  For more information on...binds cTA, resulting in a confirmation change that prevents binding to the promoter and turning transcription...

... can start your experiment? What about all the reagents you need to order? Sometimes, it feels like an... Doench also calculated the cost of consumable reagents used to make a plasmid, which came out to $115...means that the same plasmid would cost $155.25 in reagents and consumables today. Finally, this doesn’t account... the scientists designing and performing the experiments. This cost varies hugely based on a number of...assume it’s at least equal to the $155 spent on reagents. Five ways to decrease plasmid production time...plasmids with high-throughput methods that save on reagents, consumables, and time. We also handle the MTA...order to preserve time and money for their own experiments. Keep good records Oftentimes, you’ll be able...

...measure these properties, and what fascinating experiments could push these tools even further? All of these...and find further tips for using FRET in your experiments here.     What is this FRET you speak of? Förster... acceptor In order for FRET to occur several constraints must be met. The donor and acceptor fluorophores...improved dynamic range for fluorescence lifetime measurements." PloS one 10.8 (2015): e0134436. PubMed PMID...the probe. More on these biosensors later on). Experiments using intermolecular FRET probes are typically...reference standards can be used to validate FRET measurements and serve as a type of positive control. In ...questions about cellular processes. Currently, FRET experiments can probe protein-protein interactions, measure...