At the end of a process of total RNA extraction from yeast cells, when the pellets were dry, they were resuspended in RNase-free water.

In the process of extracting total RNA from yeast cells, the pellets were washed with 70% Ethanol two times.

Total RNA was extracted from yeast cells after buffering, incubation and centrifugation by phenol extraction.

4 ml of chloroform (Fischer BP1145-1) were added to buffered and incubated S. cerevisia cells in a process of total RNA extractions.

2 ml of acid phenol were added to the buffered and incubated S. cerevisia cells after transferral of the supernatant to a new tube.

After 10’ of incubation in 65 °C water bath, the samples were incubated on ice.

The yeast cells were resuspent in 3.6 ml AE Buffer and then transferred in 15 ml tubes wit 200 µl 10% SDS (w/v). 2 ml of preheated acid phenol (pH 4.3) were added and the falcon were mixed by vortex.

In a process of total RNA extraction from S. cerevisia, 2 ml of preheated acid phenol (pH 4.3) were added to yeast cells that had been collected and the pellet resuspend in 3.6 ml AE Buffer (AE=50mM NaAcetate pH 5.2, 10mM EDTA).

In a process of total RNA extraction from S. cerevisia, yeast cells were collected and the pellets were resuspended in 3.6 ml AE buffer (AE=50mM NaAcetate pH 5.2, 10mM EDTA).

In a process of total RNA extraction from S. cerevisia, the cells were collected and the pellet were resuspend in 3.6 ml AE Buffer.

MoDC were stimulated with yeast.

Monocyte-derived DCs were added to the yeast at a final concentration of 5x105 cells/ml into 96-well plates.

The melanin-free total tumor RNA is obtained from melanoma metastases; it is suitable for in vitro amplification and DC transfection.

From the CD14- PBMCs, untouched naive CD4+ T cells were isolated with a combination of magnetic sorting.

The T blasts are used to immunize MAGE-3 positive patients.

The autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24, are used for vaccination.

The patients are immunized with autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24.

The patients are immunized with autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24.

All lymphocytes (6,4 x 10e9) were frozen in 90% A-plasma and 10% GMP-grade DMSO.

The majority of monocytes (1 x 10e9) were frozen in 90% A-plasma and 10% GMP-grade DMSO.