...cells for mycoplasma (Link opens in a new window) regularly Do not over or under-grow your cells. Thaw a new...selection. It is important to monitor these cells regularly and replace the cell media. Cell death by some... or removed from the culture, check the cells regularly to confirm transgene expression. As polyclonal..., consider generating monoclonal lines from the early polyclonal populations. Sample Data Figure 1: Generation...

...properly maintain the water bath, empty and clean it regularly. Determine the amount of water needed to fill ...out after your incubation is over. Note: If you regularly use a water bath in the lab, you can leave the...previous uses. However, you will still need to regularly clean out the water bath even if you are using...

... of the BSL-2 area. BSL-2 laboratories must be clearly marked as “BSL-2.” The names and contact information...information of the laboratory manager should be clearly visible in the room and on the door. BSL-2 centrifugation...

...genome-containing virions. Phenol red is added to clearly distinguish the steps. This method will help enrich...gradient before ultracentrifugation. The steps are clearly distinguishable thanks to the use of phenol red...

...more quickly if you place the gel tray at 4 °C earlier so that it is already cold when the gel is poured...you get better separation of bands? If you have similarly sized bands that are running too close together...

... cloning is incredibly versatile and allows for nearly any piece of DNA to be placed into a backbone vector...complement to get PCR amplification. We can start similarly, taking the final 18bases of the ORF, including...

...monoclonal cell lines from the polyclonal pool as early as possible, though in theory a monoclonal line ...empirically, and that the cell population be monitored regularly for transgene expression. This protocol has been...

...you get better separation of bands? If you have similarly sized bands that are running too close together...

...this simple digest less informative. This is particularly true if you receive a plasmid from someone in...

...single reaction Ligation Independent Cloning (LIC) Scarless cloning with Type II restriction enzymes and T4...

...cells for mycoplasma (Link opens in a new window) regularly Do not over- or under-grow your cells. Thaw a ...

...storing and replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian...

...the important regions of your final plasmid, particularly the seams between the assembled parts. Tips ...

...cells for mycoplasma (Link opens in a new window) regularly Do not over or under-grow your cells. Thaw a new...

...Introduction The pipette is an essential tool for nearly anyone in molecular biology. You’ll use a pipette...

...temperature on the highest setting. Note: You should see clearly separated layers: Top Phase - Aqueous DNA phase...

...frozen. DMSO helps with denaturation of the DNA, particularly for GC-rich templates. What does each ingredient...

...37°C, 5% CO 2 overnight. Although cells should regularly be passaged in DMEM + 10% FBS with penicillin/...