Milk contains bacteria (Prokaryotic cells), Somatic Cells (Eukaryotic cells) and fats. I want to extract whole bacterial DNA from it.
I have adopted following procedure:
Milk samples were centrifuged at 5000*g for 10 mins to pellet out somatic cells. Supernatent was transferred into new tube and centrifuged at 15000*g for 15 mins to pellet out bacterial cells. Then pellets were resuspended in TE buffer or Phosphate Buffer Saline and furthered followed the same step as that for genomic DNA extraction.
Is this reliable method? I want to confirmed it because these will be used for metagenomic sequencing.
Some researchers also filtered somatic cells by using Cellulose Nitrate Membrane and then centrifuged filtrate and resuspended it in TE buffer.
Which one is better? Need your valuable suggestions on it.
Thank you in advance,
Shriram
I posted a link answer I don't know what went wrong. Briefly, I was trying to say that the thing when extracting DNA from food matrix, you need to look for a way to minimize PCR inhibitors. The paper I posted discusses about different methods doing that. Also, check your results (got using your protocol) on a Nanodrop, you will have an idea on the PCR purity. Hope it helps.
Thank you very much. The paper you suggested discusses critical aspects of extracting DNA, which is initial step of any advanced molecular technology.
Regards
The Qiagen DNeasy Blood and Tissue Kit works fine.Use it with slight modifications.(Warm the samples in ATL buffer at 60 degree with continuous vortexing)
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