Amplification of chromosomal translocation junctions from paraffin-embedded tissues of follicular lymphoma patients.
Follicular lymphoma is associated with the t(14;18) translocation, which is one of the most common chromosomal translocations in cancer. Generally, tissues from such patients are preserved as formalin-fixed and paraffin-embedded samples. Most of the time, retrieving the molecular information from such samples is hampered due to quality of preservation, extraction procedures and reaction conditions. In the present study, we isolate the chromosomal DNA from the paraffin-embedded nodal tissues of lymphoma patients and use a highly sensitive nested PCR approach to detect t(14;18) translocation. Our studies show that despite the sheared DNA obtained, appropriate modification of PCR reaction conditions can help in obtaining the desired amplifications. The DNA extraction protocol from paraffin-embedded nodal tissues and modifications in the PCR conditions are discussed. This study would contribute to the successful use of archival tissue samples in obtaining valuable information for cancer research.
Nambiar
Mridula
M
Department of Biochemistry, Indian Institute of Science, Bangalore, India.
Choudhary
Bibha
B
Rao
Clementina R
CR
Raghavan
Sathees C
SC
D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D008224:Lymphoma, Follicular / Q000175:diagnosis / Q000235:genetics*; D016612:Paraffin Embedding / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014178:Translocation, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured
D004247:DNA