...7.0. Pro-Tip The pH of this solution will drift pretty rapidly upon addition of acid or base. Add only...complete media and mix. Seed all cells in 1 CS2. Return to incubator for 48 h. Trypsinize and resuspend...CS2 into one CS5 with a total volume of 850 mL. Return to the incubator for 24 h–36 h. Proceed with transfection...new one before putting the CS5 in the incubator. Return the CS5 to the incubator for 96 h. This incubation...Distribute the media evenly between the layers. Return the CS5 to the incubator for ~72 h. Harvest cells...minutes on ice between each pulse, 50% amplitude. Return to ice between each round of sonication to avoid...

... not need to obtain more competent cells and retransform. Bacteria on an LB agar plate can be stored at...prepare your plasmid DNA. In the morning, when you retrieve your liquid bacterial culture, take 500 μL of ...

...resolution (the ability to distinguish between two discrete objects). Figure 1 depicts an image of a compound... sample information so that you or others can interpret your images in the future. Conclusion Like any...

...results in cell populations that are more likely to retain stable transgene expression. Other methods of generating...well in sparse/individual cultures due to lack of secreted growth factors, so not every cell type will be...

...position 15-19. Avoid targeting introns. Avoid stretches of 4 or more nucleotide repeats, especially repeated...Incubate for 20-30 minutes at room temperature. g. Retrieve HEK-293T cells from incubator. The cells should...

...end, and the 5’ end of the primer usually has stretches of several nucleotides. Also, both of the 3’ ends...

...cloning can be used anytime you need to add a short stretch of DNA to a plasmid, such as: Changing a Multiple...

...cloning scars"). A “stuffer” sequence allows for electrophoretic separation of linearized vector from the reaction...

...Modification by Annealed Oligo Cloning Add a short stretch of DNA to a plasmid Gibson Assembly Combine overlapping...

...first. Pro-Tip The pH of this solution will drift pretty rapidly upon addition of acid or base. Add only...

...indicates the various controls: Control Ligase Interpretation Uncut vector - Checks viability of competent...

...dropwise. Cap the flasks and swirl 5–10 times to mix. Return the flask to the incubator. Section 3: BCD Feed...

...details on doing diagnostic digests and how to interpret them please see the Diagnostic Digest page. Purifying...

...manufacturer to make sure this is true in your case. Retrieve your molten agar mix from the autoclave. Pro-Tip...

...little chance for mis-priming, so you can use a pretty wide range of annealing temperatures, but you may...

...background staining. Pro-Tip A large shift in the electrophoretic mobility of the bands (sample AR0016 in Figure...