Sequence analysis of amplified t(14;18) chromosomal breakpoints in B-cell lymphomas.
We have explored different strategies for sequencing of major breakpoint (mbr) junctional regions in t(14;18) chromosomal translocations--the most frequent chromosomal abnormality observed in B-cell lymphomas. We demonstrate that coupling of the preparation of single-stranded DNA by asymmetric polymerase chain reaction (PCR) and direct sequencing is the method of choice for the rapid and precise determination of clone-specific bcl-2/JH fusion gene sequences. The rapidity, relative ease, and accuracy of the technique, described for the nucleotide sequence analysis of mbr t(14;18) breakpoints, permits the analysis of a relatively large number of samples and should be considered as part of the clinical evaluation of lymphoma patients. Furthermore, by providing sequence information of clone-specific DNA regions, the procedure should reduce the risk of false-positive results from PCR.
Eick
S
S
Abteilung Hämatologie/Onkologie, Zentrum Innere Medizin, Klinikum der Universität Göttingen, F.R.G.
Krieger
G
G
Bolz
I
I
Kneba
M
M
D001483:Base Sequence; D002883:Chromosomes, Human, Pair 14 / Q000737:chemistry*; D002887:Chromosomes, Human, Pair 18 / Q000737:chemistry*; D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D016393:Lymphoma, B-Cell / Q000235:genetics*; D008969:Molecular Sequence Data; D009857:Oncogenes / Q000235:genetics; D016133:Polymerase Chain Reaction; D014178:Translocation, Genetic / Q000235:genetics*
D004273:DNA, Neoplasm