Rapid cloning of rearranged immunoglobulin heavy chain genes from human B-cell lines using anchored polymerase chain reaction.
A general method to obtain the variable region DNA sequence of the immunoglobulin heavy chain using anchored polymerase chain reaction is described. Based on this DNA sequence, clone-specific oligonucleotides were designed to anneal to the complementarity determining regions. These were used to identify the original B-cell clone in serial dilutions of polyclonal lymph node DNA with high specificity and sensitivity. This method should be useful for studying minimal residual disease in B-cell neoplasia.
Ratech
H
H
Department of Pathology, College of Physicians & Surgeons, Columbia University, New York, NY.
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods
D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D004247:DNA