...Protocol, tips, and FAQ for how to perform a restriction digest of plasmid DNA...more enzyme is advisable. Reactions are often performed with 0.2-0.5 µL of enzyme because it is difficult...

...ll express your gRNA and Cas9 protein, which will form a ribonucleoprotein complex through interactions...effector, composed of several separate Cas enzymes that form a complex called Cascade (see Cas3 for more details...with Cas3 Rather than initiating DNA cleavage to form a break, Cas3 initiates single-strand DNA degradation...dCas9 Nuclease dead Cas9, an enzymatically inactive form of Cas9; can bind, but cannot cleave DNA DSB D ouble...Cas9 undergoes a second conformational change upon target binding. In this formation, its nuclease domains...detection platform. Based on this non-specific cleavage, a Cas13a-based molecular detection platform, termed...positively-charged grooves on Cas9. gRNA binding induces a conformational change in Cas9, shifting it into an active,...

...containers Reagents Liquid for pipetting Background Information Anatomy of a Pipette This front view photo of...the Volume on the Pipette Although each pipette performs the same function, the numbers are read differently...

...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and...Watch the protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well...

...Follow this protocol to perform digital droplet PCR (ddPCR) for lentivirus vector titration....Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence Titration...

...Restriction Digest of Plasmid DNA Background Information During several different stages of molecular ...

... capabilities. Taking into consideration the information above, primers should generally have the following...

...PSAM is coupled with an ion pore domain (IPD) to form ligand-gated ion channels (LGICs) that can either...

...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture...

...Plasmid DNA from Bacterial Culture Background Information Plasmids can carry one or more antibiotic resistance...

... (am): Observe fluorescence, harvest cells, or perform your experiment Equipment Class II, Type A2 Biological...

...You can purchase master mix or make your own. Transform bacteria with the DNA and screen for the correct...

... D E Sample 1 Sample 4 F E Sample 2 Sample 5 F Perform data analysis using the instrument’s software. ...

...specific products or equipment. Inclusion of this information is solely for transparency intended to support...

...Follow this protocol to perform digital droplet PCR (ddPCR) for purified AAV titration....

...serotypes are harvested at 96–120 h. Optional: perform a media exchange at ~18 h post transfection. Changing...

...experiments being performed, and your biosafety office can provide more information on your institution's...plasmids utilize a hybrid LTR promoter (more information on this below). Additional or specialized promoters...different viruses and thus use slightly different isoforms of these packaging components. Therefore, lentivirus... a range of species and cell types. For more information, see the Cronin, et al. article on different ...the lentiviral genome and harbor the genetic information encoded by that genome. Many lentiviral transfer...generate more useful restriction sites. For more information about cloning and changing a MCS, visit Addgene's...recombine. Visit Invitrogen's website for more information on the Gateway® cloning system. What safety concerns...

...plasmid. Please see our Biosafety guide for more information on working with adenovirus. Overview of Adenoviral...restriction enzyme PmeI. This construct is then transformed into AdEasier-1 cells , which are BJ5183 E. coli... into the adenoviral plasmid. One can also co-transform standard BJ5183 with supercoiled pAdEasy™ and ...E1, E4, E2a and VA for replication. For more information about AAV, read our AAV guide . Can I make a ...each round of amplification. Where can I find information about using the AdEasy™ System? The laboratory...

...your library from Addgene, check the library information to see if it should be amplified before you conduct...in the selection NEGATIVE screen: Apply library Perform Next Generation Sequencing (NGS) on a control sample...library Apply selection; most cells will survive Perform NGS on the surviving cells Compare NGS results ... Company Resources Science Exchange Scientist (formally Assay Depot) Resources Find additional resources...amplification protocol. Read our CRISPR guide for more information on genome-wide screening with CRISPR/Cas9 . Browse...

...products for packaging (i.e., Gag, Pol, and Env), the isoforms of these proteins, as well as the viral LTRs, ... Addgene . Please see our Biosafety page for information on working with retrovirus. Packaging using 293T...different viruses and thus use slightly different isoforms of these packaging components. Therefore, lentiviruses...plasmids may be used interchangeably. For more information about lentivirus, please read our lentivirus ...