I did this by first making stock solutions of 1M Tris-Cl (pH 8.0) and 0.5M EDTA, which are the normal working concentrations for these.

Then I prepared a  1L buffer by adding

20ml of 1M Tris-Cl (final concentration of 20mM Tris-Cl in buffer)

2ml of 0.5M EDTA (final concentration of 2mM EDTA in buffer)

12ml Triton® X-100 (final concentration 1.2% Triton® X-100 in buffer).

Before I added the lysis buffer to the sample, I added 20mg of lysozyme to 1ml of 

the buffer, so you will need 100mg lysozyme for your 5ml of buffer.

I had really good results after my extraction. Hope this is helpful.

Grace, did you add salts at any point? Sorry, am getting confused where the precipitation step occurs in my protocol. I am using the same mix but with Tris-HCl, EDTA, Triton-X and lysozyme. I noticed the original question used sodium EDTA?

Hi Jessica,

I used sodium EDTA as well, sorry I omitted the sodium in my previous answer ...Are you using the Qiagen dNeasy kit for your extraction? The extraction is quite easy and straightforward with their protocol.

Could you explain the challenge you are having with your extraction if your protocol is different...maybe we could figure it out.

Hi Grace,

When you say you made up the stock solutions in buffer e.g. final concentration of 20mM Tris-Cl in buffer what do you add to make the final volume 1L? Is it PBS or distilled water? 

Thanks,

Thanks!

Hi Tayhla,

I prepared it with sterile distilled water. Sorry for the late reply, I don't check this page frequently. All the best in your research.

If someone needs to use above mentioned stock conc. of EDTA 0.5M to get final conc. of 2mM in 1L solution then you need to use 4 ml instead.