I am trying to isolate bacterial genomic DNA from lactobacillus which is incubated +4 degree (around one week), and in a tube (inside liquid media). But I could not get a high DNA concentration from them. I dont think it is related source but I am confused. Can you help me anyone? Thanks in advance.
Hi Serife,
The fresh bacterial cultures are generally recommended to isolate genomic DNA because the bacteria are not stressed with the depleting nutrients and the accumulation of the waste products in the broth. To get more concentration you can select the mid to late exponential phase bacterial culture. Moreover, you are isolating genomic DNA from Lactobacillus , the cells in this case can be treated with lysozyme in the lysis step so that more DNA can be extracted.
It depends to some degree on the intended end use of your genomic DNA preparation. For example, if it is for direct hybridisation, you will want ample quantities of unsheared gDNA. But if it for use as a template involving an amplification step, that is generally more forgiving. The issue may be the gDNA isolation protocol, but without knowing the specific details of the protocol being used or the end use of the gDNA, it is difficult to identify where in the process the problem is occurring.
I don't think it matters much if your bacterial culture is fresh or stored in freezer. Practically, in lab environments, DNA is isolated from the frozen samples and the resulting DNA quality is good enough for the general usage. However, DNA extraction method can influence the amount and quality of the extracted DNA a lot. Thus, as Ronan O'Toole said, troubleshoot your extraction protocol.
Follow This Link Please:
https://www.encyclopedia.com/science-and-technology/biology-and-genetics/genetics-and-genetic-engineering/dna-isolation-methods
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