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DNA PURIFICATION QUESTIONS:
Why is it important to remove as much as possible of the growth medium from the cell pellet during cell harvesting?
Alice and Frankie are lab partners in a micro lab performing DNA extraction, each person using a different bacterial species. Both of them spun 1.8 ml of their overnight broth cultures in a microcentrifuge, and it turned out that Aliceâs tube had a substantially larger pellet than Frankieâs. What could Frankie do to increase the amount of cell pellet necessary for subsequent extraction steps?
I am performing a gel extraction to purify DNA after a doubledigest with EcoRI and BamHI. After the gel extraction I need tocomplete a ligation step before bacterial transformation. Theproblem I am having is that the DNA after the gel extraction havesome kind of chemical contaminant that won't allow me to quantifythe DNA using the Nanodrop spectrophotometer. I am getting very low260/230 ratios.
Has anyone had some issues with this?
I am using the Qiagen Qiaquick Gel Extraction Kit.
