I am trying to do DNA extraction from two strains of gram-positive sulfate reducing bacteria which form MeS precipitates (Zn and Fe). I am having some problems with DNA yields, they are two low! as I need them for Whole Genome Sequencing. I've tryed several protocols like phenol:cholorophorm:isoamilic alcohol, and CTAB. Does anyone have suggestions??
Thanks a lot!
Have you tried commercial kits from Sigma or Invitrogen - they work very well for my lab
Have you tried commercial kits from Sigma or Invitrogen - they work very well for my lab
though it is time consuming method ,manual method using phenol chloroform isoamylalcohol(24:24:1) for midlog phase culture is the best method to isolate DNA from gram positive bacteria
Check the following point:
All the best for your good results.......
An economic and effective DNA extraction method we use for our Listeria monocytogenes, a member of Gram-positive bacteria, is the bead method: Please find the method in this article: http://www.sciencedirect.com/science/article/pii/S0167701205001077
Or
http://www.ncbi.nlm.nih.gov/pubmed/15935495
It is every time challenge - but anyway you need to remove cell wall first - so try lysozyme overnight - and after that proceed with any column purification protocol or traditional phenol chloroform extraction after proteinase digestion.
You also can try to lyse huge amount of bacteria in exponential phase of propagation
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