Feasibility of archival non-buffered formalin-fixed and paraffin-embedded tissues for PCR amplification: an analysis of resected gastric carcinoma.

摘要: Although several factors affecting the sensitivity of polymerase chain reaction (PCR) amplification from formalin-fixed tissues have been investigated mostly by experiments, feasibility archival formalin-fixed, paraffin-embedded tissue samples stored in pathology departments for PCR has rarely examined directly. Thus, 74 unbuffered 10% with primers producing a 190 b.p. DNA segment p53 exon 5 was investigated. Fixation time critical factor influencing amplification. All (6/6) fixed only 1 day, 44% (7/16) 2-3 days and 14% (4/28) 4-6 showed successful amplification, while no obtained 7 or more. The peak size extracted decreased as fixation became longer. Experiments using xenografted tumor various times longer permissible time; up to 9 fixation, decreasing amounts products were 12 paraffin seemed be minor since all day samples, including those that had embedded years, resulted efficient amplified segments, however, could another because even less 345 compared 167 262 DNA. Additionally, point mutation detected non-isotopic method, temperature gradient gel electrophoresis. In conclusion, immediately constantly available approximately 200 suggesting surgical specimens should subjected cutting embedding just after subsequent use