Chimeric molecules created by gene amplification interfere with the analysis of somatic hypermutation of murine immunoglobulin genes

Gene. 1994 May 16;142(2):279-83. doi: 10.1016/0378-1119(94)90275-5.


We used the polymerase chain reaction (PCR) to amplify genes encoding murine immunoglobulin (Ig) lambda light-chain variable (V) regions, using DNA isolated from populations of germinal center B-cells, to study somatic hypermutation at this locus. Sequence analysis revealed that 30% of the amplified products were chimeric molecules consisting of segments of the V lambda 1 and V lambda 2 genes. Furthermore, an amplification- and cloning-associated artifact exchanged sequences between mutational variants of V lambda 1 genes. These PCR artifacts interfere with the analysis of somatic hypermutation of Ig genes. An alternative method that avoids these artifacts is suggested which involves the amplification of individual V lambda genes from single cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Artifacts*
  • B-Lymphocytes / immunology
  • Base Sequence
  • Cloning, Molecular
  • DNA Mutational Analysis
  • DNA, Recombinant / chemistry*
  • Genes, Immunoglobulin / genetics*
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin lambda-Chains / genetics
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Mutation / genetics*
  • Polymerase Chain Reaction*


  • DNA, Recombinant
  • Immunoglobulin Variable Region
  • Immunoglobulin lambda-Chains