... Restriction Cloning and Fast Cloning Requirement Restriction Cloning Fast Cloning Choose Restriction... Transformation + + The FastCloning process in brief In FastCloning, you PCR both the vector and ...and ligated in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and vector...most importantly, excessive time requirements. FastCloning is a simpler yet reliable cloning technique that...understanding of the step by step procedure of FastCloning, here are some tips for high efficiency cloning...of the colonies are correct. Good luck with this fast and easy technique! Many thanks to our guest blogger...PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and...

...effects can mask weaker ones. Thus, the need for a faster and higher-throughput in vitro approach. Using ...constructed as well. They called this library the CTD2 FASMIC Plasmid Collection for “Functional Annotation of...

...high levels of correct assemblies) translate to faster assembly workflows that require less screening ...

.... https://doi.org/10.1038/nmeth.3993  Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X,...doi.org/10.1038/s41586-019-0908-x  Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani.... https://doi.org/10.1016/j.cell.2014.02.001  Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ...

...Thakore PI, Moreb EA, Rivera RM, Madhavan S, Pan X, Ran FA, Yan WX, Asokan A, Zhang F, Duan D, Gersbach CA. ...Widrick JJ, Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe LH, Church GM, Wagers...25692716. PubMed Central PMCID: PMC4335351. 9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ...

...such as 2-fluoroadenine (2-FA), organisms with functional APT will convert 2-FA to cytotoxic nucleotides...nucleotides, resulting in death. In the presence of 2-FA, potent gene silencing of APT is required for survival...Darren Gilmour at the University of Zurich. He is fascinated by how organisms exquisitely and robustly self-organize...

...doi.org/10.1177/1948550619882032 Chrousos GP, Mentis A-FA (2020) Imposter syndrome threatens diversity. Science.... https://doi.org/10.1126/science.aba8039 Cordova FA (2016) Embrace uncertainty. Science 351:994–994 ....

...AR.  Nature. 2013 Jan; 493, 429–432. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X,...

....   References and resources References Al-Allaf FA, Tolmachov OE, Zambetti LP, Tchetchelnitski V, Mehmet... containing the recombined product tend to grow faster, so we recommend picking the smaller colonies which...

... https://doi.org/10.1016/j.molcel.2014.08.002 Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, Trevino...breaks in tens of minutes – an order of magnitude faster than HDR. NHEJ is consequently the principle means...

...complex with guide RNA and target DNA. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani...

...556:57–63 . https://doi.org/10.1038/nature26155 Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ...

...Genome engineering using the CRISPR-Cas9 system. Ran FA, et al. Nature Protocols Nov;8(11):2281-308; (2013...

...that better human health.     References Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X,...Like everyone else in the world, I (Wenning) was fascinated by CRISPR and rushed to test it. I still remember...

...https://doi.org/10.1016/s0764-4469(00)01250-6 Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F ...

...The idea was to evolve a TadA-7.10 version with a faster deamination rate so that deamination can happen...PANCE) selection systems to engineer better and faster ABEs (Fig. 2). These systems allow for rapid continuous...speed dictates which phage variants can propagate faster than the rate of dilution (passage from the infected...activity and editing speed. Introducing ABE8e - a fast and flexible adenine base editor Lower-stringency..., they identified TadA-8e, which edits 590-fold faster than TadA-7.10. It is compatible with all Cas9 ...resulting base editor was named ABE8e. ABE8e is not only fast, it can also be multiplexed: when tasked to correct...functional domains speed up the development towards faster and more precise base editors capable of editing...

...check out our blog post on choosing a Cas9 enzyme. Fast facts – PAM requirement: NGG and an expanding number...has anti-viral application potential as a result. Fast facts – PAM requirement: AAG or TTC  Best for: very...optimize it for therapeutic and laboratory usage. Fast facts – PAM requirement: TTN or TTTN. Best for:...variants such as single nucleotide polymorphisms. Fast facts – PAM requirement: none. Best for: high fidelity...information on Cas13, check out our blog on RNA editing. Fast facts – PAM requirement: protospacer flanking sequence...11 in our blog post on its development and use. Fast facts – PAM requirement: protospacer flanking sequence...targeting single stranded RNA.   Table 1: Fast facts on Cas family proteins and genome editing ...

... below use the following optogenetic modules: VVDfast homodimerizes under blue light iLID-SspB heterodimerizes...limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule severing. A) Schematic of...of Opto-katanin constructs and activity. EB3N-VVDfast-iLID (anchor) and SspB-mCherry-p60 (katanin) are... of two separate blue light-sensitive systems, VVDfast to control recruitment of an “anchor” to microtubules...anchor. So, you’ll need to express both the EB3N-VVDfast-iLID anchor module and the SspB-mCherry-p60 katanin...controlling cell dynamics. Need a refresher on VVDfast, iLID, LOV, CRY2-CIB, and other optogenetic modules...

...Verkhusha Multiple FAST Fluorescence-activating and absorption shifting tag (FAST) for use with green-yellow...superpositive GFP Split-superpositive GFP reassembly is a fast, efficient, and robust method for detecting protein-protein...mScarlet Split-wrmScarlet and split-sfGFP: tools for faster, easier fluorescent labeling of endogenous proteins...requires some time for chromophore maturation, splitFAST displays rapid and reversible complementation....