We narrowed to 357 results for: Plat;

...Keisuke Kaji Jonathan Katz Nathan Lawson Kathrin Plath Miguel Ramalho-Santos Derrick Rossi Michael Ruppert...

...Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed...

...Esposito Lab for use with the Gateway® Cloning Platform (Life Technologies™) to permit construction of...

...Jaramillo Lab Riboregulation Keasling Lab BglBrick Platform , Terpenoid Production , and Propionate Induction...

... in the collection have been developed using a platform (Link opens in a new window) based on yeast display...

...binding site (PBS) and the desired edits on an RT template. During prime editing, the pegRNA directs the ...

...set of plasmids for building homologous repair templates that incorporate a self-excising drug selection...

...promoter, forms blue colonies on LB/Amp/Xgal/IPTG plate pAdx-CMV-copGFP 73346 Expresses copGFP from the ...

...bind adenenine with lower specificity). With the template for studying this type of technology already laid...

...isolating genomic DNA. This DNA is then used as a template to amplify a portion of the intended lentiviral...

...coronaviruses. Addgene assisted this effort by providing a platform for the distribution of plasmid-based tools that...

.... protoSpaceJAM protoSpaceJAM is an all-around platform for CRISPR knock-in design at scale. It uses an...

...transfection into plates containing selection agent at different densities, then dispose of the plates that do ...) to transition from a 48-well cell plate to a 96-well assay plate. We generally freeze at least 2 ampules...cloning rings, the remainder of the colonies on the plate dried out and died. The alternative to cloning rings... rings, dilution cloning into 96-well plates, is extremely time- and resource-consumptive, since only ...cell can give rise to single clones, and thus many plates must be examined for single clones and then handled...12-48 individual colonies by (carefully) holding plates up to the light, where they appear as opaque dots... achieved by splitting transfected cells into 3 plates at different densities for selection). A 2% agarose...

...column, plate Mini, midi, maxi, mega, giga $ Yes —beads, columns, plates, buffers 96-well plates, 8-well...Beads, column, plate Mini, midi, maxi $ Yes — beads, columns, plates, buffers 96-well plates, bead kits, ... Column, plate Mini, midi, maxi, mega, giga $$ Yes — columns, plates, buffers 96-well plates, automation-friendly... column, plate Mini, midi, maxi, giga $ Yes — beads, columns, plates, buffers 96-well plates Not mentioned...Millipore Sigma Column, plate Mini, midi, maxi, mega, giga $$ No 96-well plates Some Endotoxin-free, BAC...Promega Beads, column, plate Mini, midi, maxi $$ Some — buffers 96-well plates, automation-friendly In...Fisher Beads, column, plate Mini, midi, maxi, mega, giga $ Some — buffers 96-well plates, automation-friendly...

...modifications exist in the template DNA. There are several unique HDR mechanisms, but all templated-repair pathways...along the way. HDR uses homologous sequence as a template for repair and is thus the more accurate mechanism...process is generally error-free provided the DNA template used for repair is identical to the original DNA...one strand of the homologous DNA donor and can template repair off the other strand; this results in the...mechanism that notably does not require a donor template and thus won’t be covered in detail here. Classical...pathway, the 3’ ends invade an intact homologous template to serve as a primer for DNA repair synthesis,...portion of the invasive strand is displaced from the template and returned to the processed end of the non-invading...

...requires a template of high homology to the endogenous target, yet to facilitate the repair, the template should...protocol that amplifies the entire plasmid template. The parent template is removed using a methylation-dependent...resistant to Cas9 cleavage. If a plasmid contains the template, site-directed mutagenesis can be used to mutate...plasmids. PCR primers (green) amplify the plasmid template (blue), and introduce a restriction site “A*” ...the targeted region. Upon PCR amplification, the template is removed by DpnI restriction digestion leaving...circle rather than a linear product (see figure). Template A high purity plasmid prep significantly increases...you may want to try different concentrations of template (e.g. 0.1-1.0 ng/μl). Smaller plasmids (~3 kb)...

... Single-stranded DNA repair templates Short single-stranded DNA templates have been shown to improve the...final ssDNA template. Figure 2: Overview of ivTRT to generate an ssDNA repair template. Although...from low efficiency when a standard dsDNA repair template is used. The original design for creating a conditional...targeting two intronic regions and two repair templates in order to insert two loxP sites. Since two double-strand...ameliorate this problem. Briefly, you design your donor template to include a T7 promoter upstream of the left ...vitro transcription, and use the IVT product as a template for reverse transcription to create your ssDNA...tested by Quadros et al. When delivering the ssDNA template and CRISPR machinery, Quadros et al. recommend...

...target in the HR template, you usually would need to make mutations of the HR template to avoid this donor... HR template an issue? A8: Yes. If you have intact protospacer+PAM sequence within the HR template, it...homologous recombination (HR) Q7: Can you summarize HR template vector design guidelines? A7: A few notes below...you should consider using ssDNA oligo as donor template, it's usually more efficient than plasmid donor...µl of 10 µM oligo stock for each well on 24-well plate, co-deliver into the cells with the Cas9/sgRNAs ...with Cas9/sgRNAs vector, for one well in 24-well plate. A good example/reference is Ran et al. (2013). ... good way is to mutate the PAM 'NGG' in the HR template, like change it to 'NGT', or 'NGA', or 'NGC' (...

...and plate transformants for isolated colonies (150-200 per plate) across 20 fresh selective plates. Step...gently down onto a plate of colonies. Mark the orientation of each membrane on its plate before moving on...on. Then peel each membrane off the plate (saving the plate so you can regrow the lifted colonies overnight...outcompeted by others in liquid culture, dilute and plate the complete library of transformants for single...single colonies (this usually requires 10-20 plates) (see box below). This initial transformation should be...over and place it colony-side up onto a fresh LB plate containing an appropriate inducing agent (0.4 mM...protein induction is done, peel the membranes off the plates and place each onto a lysis solution-soaked filter...

...the template. The opposite is true if the sense strand is nicked for template formation: template will...low primer-to-template ratio is used to promote annealing of one primer to each template. The PCR product...Error-prone PCR: This method amplifies the wild-type template with a “sloppy” version of PCR that results in...). It uses a uracil labeled wild-type ssDNA as template for a site-directed mutagenesis PCR that uses ...strand (step 2 and 3). Then the uracil-labeled template strand is degraded by uracil DNA glycosylase (...present on both strands of the plasmid. The ssDNA template is generated by transforming a backbone phagemid...days to generate the phage-derived uracil labeled template, this method requires further DNA shuffling to...