The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. III. Extraction and purification of RNA and application to slot-blot hybridization analysis.
RNA was extracted from tissues processed by a new fixation and paraffin-embedding method (the AMeX method) and examined by Northern blot analysis and slot-blot analysis. The RNA extraction method for AMeX-processed tissue sections after the deparaffinization step was the same as that for fresh materials. The total amount of cellular RNA extracted from AMeX-processed mouse liver tissue was slightly less than that extracted from fresh tissue. In tissues of malignant lymphoma, the total amount of cellular RNA extracted from 25 sections each 20 microns thick was about 1.6-1.8 micrograms/mm2, regardless of the histological subtype and period of storage. The extracted RNA was moderately degraded, and usually could not be used for Northern blot hybridization analysis. The intensity of ethidium bromide staining and the hybridization signals of RNA extracted from AMeX-processed tissues were usually reduced in comparison with RNA from fresh material, but specific signals could be detected by slot-blot hybridization analysis. We have demonstrated previously that the AMeX method preserves high-molecular-weight DNA and various antigens. Since the present study showed that information on mRNA can be obtained from AMeX-processed tissue, the versatility and usefulness of this method were further proven.
Sato
Y
Y
Pathology Division, National Cancer Center Research Institute, Tokyo, Japan.
Mukai
K
K
Furuya
S
S
Shimosato
Y
Y
D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D008099:Liver / Q000737:chemistry; D008223:Lymphoma / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008867:Microtomy; D009693:Nucleic Acid Hybridization; D010232:Paraffin; D012313:RNA / Q000302:isolation & purification*; D014021:Tissue Preservation
D012313:RNA; D010232:Paraffin