Dear all:

I´m trying to extract DNA from agave silages samples, our samples have been concentrated and purified by percoll density gradients. However, DNA quantity and quality are low.

Briefly:

1.- Lysis: samples were placed in 500 uL of lysis buffer (300mM tris HCL, 30mM EDTA, 800mM guanidium HCL, 0.5% triton, pH10). 0.300 g of glass bead 100 M. Freezing-thawing, and heat 1 h at 50°C. And vortex max speed 10 min.

2.- Purification: Cloroform-isoamilic alcohol 24:1 twice

3.- Precipitation: linear acrilamide, PEG 6000 30% + 1.2 NaCl +, incubated room temp for 2 h

4.- Two alcohol 80% washes

5.- 50 ul TE buffer, incubated 2 h, 42°C

Mean DNA Quantification 20-30 ug/ul

Any advice will be helpfull