I am currently working on DNA purification from peptides (protein) by using silicon magnetic beads. I can not get results. I think I am making some mistakes in my protocol, it will be very appreciated if someone can help me.

My experimental solutions are

1: Lysis byffer

a. Tris-HCl (Tris + HCl)

b. pH 7.6

2. Binding solutyion

a. Tris-HCl

b. 1% SDS

c. NaCl

Process: I am taking 10ml binding solution and add 40% (13.33ml) isopropanol, 10ml protein and 1ml magnetic beads (after washing). I mixed them well and then put it on shaker for 20 minutes at 100rpm to bind and mix.

Then I seperate the white supernatatnt through magnet and lebel it and washed the remaining magnetic pallet with washing buffer ( Binding sol+ isopropanol+ ddwater), and then added Tris-HCl and put it in hot water (60C) for 10 minutes.

After that I take the supernattant and labeled (washed solution).

I run the SDS by following sequence

a. Orignal protein

b. Diluted protein (700% protein + 30% dd water)

c. Supernatant after 20 min shaker

d. Washing solution used for beads after 20 minutes shaker

e. Supernatant after elution (60C) of 10 minutes (lowe solution).

I run these samples on SDS but did not got bands for C, D and E. I only fot results for a and B. which were used as a control.

If anyone is working on magnetic beads for DNA purifiaction, please help me in this process. I will be very thankful for your kindness.