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... of powder in 100 mL of deionized water. While stirring, slowly add hydrochloric acid until the solution...DNA tube. Incubate the mixture 12–15 min at RT. During the incubation, add 10 mL of DMEM Complete to a... pellet any packaging cells that were collected during harvesting. Filter supernatant through a 0.45 μm...

...knobs on the side of the microscope. You may be wondering why there are multiple objective lenses. Each ...with the lighting, use the coarse focus knob to bring your sample into focus. Once your sample is in focus...easily see your sample. Then use the focus knobs to bring the sample into focus. Pro-Tip As you increase your...

...reference AAV of known titer, 100837-AAV1, and by measuring the titer of samples obtained from academic viral...transparent film. Centrifuge at 3,000 rpm for 2 min to bring the sample to the bottom of the tube. Run the following...amount of background signal that is most evident during initial PCR cycles. This background signal must...

... the sample is determined. Sharing speeds science. We believe that sharing the full details of our protocols...and ImageJ software. The sample is separated by denaturing polyacrylamide gel electrophoresis alongside ...

...SDS-PAGE gel, and immunoblotting. Sharing speeds science. We believe that sharing the full details of our protocols...buffer is suitable for most proteins but more stringent buffers and a sonication step may be required ...

...feet from potential spills. If wearing open-toed shoes, cloth coverings or “booties” can be used to protect...

...Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Introduction... you have single colonies, you can proceed to Recovering Plasmid DNA or use the individual colonies for...

...with the razor blade. This is especially important during the DNA purification step, as many kits cannot ...used to determine how much of each buffer to add during the DNA isolation step. Finally, you will want ...

... cloning methods. In traditional cloning, base-pairing in the short overlapping regions (usually 4 bp)...annealed but nicked vector product is then repaired during the replication cycle. Empty vectors for LIC typically...

...Guidelines BSL-1 Guidelines Before You Work Right after entering the lab, put on the proper personal protective...concentration of 10% bleach for 30 minutes before pouring down the drain. Solid BSL-2 waste can be collected...

...cells, target proteins, etc. Sharing speeds science. We believe that sharing the full details of our protocols...

...target to show a dose response. Sharing speeds science. We believe that sharing the full details of our protocols...

...immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples, primers/probe mixes, and...samples on ice or pre-chilled 96-well freezer blocks during use. To reduce the risk of contaminating reagents...

...inosine) instead of a regular nucleotide for broader pairing capabilities. Taking into consideration the information...

...You may also like... Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Background...

...oligos. This is an option that can be added when ordering them or can be performed enzymatically later. ...

...additional charge. What primers were used by Addgene during quality control? Addgene lists the primers used...

...your container. Make sure that any spills produced during the protocol are cleaned up. Discard the weighing...

...bath is a piece of laboratory equipment that helps bring your materials to a particular temperature, catalyze...

...Introduction Restriction enzymes are naturally occurring bacterial endonucleases that recognize a large...