I am using the NEB Monarch Genomic DNA Purification Kit to extract gDNA from BL21(DE3) cells. The protocol says to use a maximum of 2x10^9 cells but does not mention a recommended culture volume or pellet mass to use. I have read varying information on the amount of E. coli cells per mL of overnight culture, so I am wondering what the recommended volume of culture/pellet mass for gDNA extraction of BL21(DE3) is?
Hi August Brookwell
Genomic DNA Extraction Basics Solution-based methods for DNA purification rely on precipitation and centrifugation steps to separate the genomic DNA in the cell lysate from other cellular materials. These methods use either organic extraction or “salting out” to separate soluble DNA from cellular proteins.
Follow this link and gain huge information..
https://www.neb-online.de/en/monarch-nucleic-acid-purification-kits-2/new-monarch-genomic-dna-purification-kit/
Let reach the OD of the culture between 0.8 to 1. Overnight culture can be use for DNA isolation. However, I didn't try above mention kit. I used miniprep kit of QIAGEN.
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