Dear Dr Ana Paula Palacios,

I found an article, in which researchers investigated the DNA extraction of other microorganisms strains using SDS. These strains were respectively; Enterococcus faecalis, Corynebacterium diphtheriae, Escherichia coli and Staphylococcus aureus. Indeed, they used 2 procedures with different incubation times, I have summarized you the two procedures as follow:

Procedure A:

Bacterial pellets were resuspended in 0.2 ml of digestion buffer [50 mM Tris-HC1 (pH 8.5), 1 mM EDTA, 0.5% SDS, 200 ug/ ml proteinase K] and incubated for 3 hr at 55°C with agitation (Thermomixer, Eppendorf). After heat inactivation of the proteinase K for 10 min at 95°C, the tubes were cooled to 4°C and centrifuged for 10 min at 12,000g. Ten microliters of the supernatant was used directly for PCR amplification while adding 2% Tween 20 (final concentration) to the amplification mix.

Procedure B:

Bacterial pellets were resuspended in 0.2 ml of Triton X-100 buffer [10 mM Tris- HC1 (pH 8.0), 1 mM EDTA, 1% Triton X-100] and incubated for 30 min at 95°C with agitation. The tubes were then cooled to 4°C and centrifuged for 10 min at 12,000 and 10 ul of the supernatant was used directly for amplification.

You will also find in this attached, the article source with all details.

Best wishes,

Sabri

Dr. Mohamed Sabri Bensaad

I'm really grateful for answering my question. I will try to do these procedures and read more concerning extraction. Thanks for attaching the article.

Best wishes

Ana Paula

Your welcome Dr Ana Paula Palacios.

All the best for you.