Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue
Fig 4
100 mg of frozen adipose tissue was fixed in 0.5% paraformaldehyde and then homogenized using the Ultraturrax method.
It was then sheared for 40 cycles (30 seconds ON and 30 seconds OFF), after which a sample of 50 μl of the homogenized material was taken, and the chromatin was de-crosslinked using either the fast Chelex-100 method or a moderate temperature for 5 hours plus PK treatment. Fig 4 shows the de-crosslinking step at a moderate temperature for 5 hours and then a PK step improves the quantity of DNA with respect to the method based on the use of Chelex-100 (n = 6) Total DNA was extracted and quantified by nanodrop. Data are given as means with error bars.