Isolation of nucleic acid from paraffin embedded tissue for PCR amplification and sequencing of TcR V beta genes.
PCR analysis of DNA and RNA from fresh and frozen tissue is now a routine practice in most laboratories. Furthermore DNA extracted from paraffin embedded tissue is routinely used for clonality studies and for HLA typing. However the use of RNA from paraffin embedded tissue is more problematical due to degradation and only short sequences are suitable for amplification. In this report we examine the suitability of extracted nucleic acid from paraffin embedded tissue for the PCR amplification of TcR V beta genes and sequencing of resultant PCR products.
Hodges
E
E
Wessex Immunology Service, Southampton University Hospital NHS Trust, U.K.
Smith
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D010232:Paraffin; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D013997:Time Factors; D014021:Tissue Preservation
D016693:Receptors, Antigen, T-Cell, alpha-beta; D012313:RNA; D010232:Paraffin; D004247:DNA