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Modification of IgH PCR clonal analysis by the addition of sucrose and cresol red directly to PCR reaction mixes.
  1. E Hodges,
  2. S M Boddy,
  3. S Thomas,
  4. J L Smith
  1. Wessex Immunology Service, Southampton University Hospitals NHS Trust, United Kingdom.

    Abstract

    Diagnostic immunoglobulin (Ig) polymerase chain reaction (PCR) clonality analyses need to be simple, reproducible, and rapid. Sucrose and cresol red (gel loading buffer reagents) were added to a routine IgH PCR reaction mix to obviate the need for adding gel loading buffer separately after PCR amplification. Not only did this decrease the time spent after PCR analysis but also gave similar or enhanced IgH PCR product intensity compared with normal IgH PCR conditions on polyacrylamide gel electrophoresis. This procedure was easily adapted to routine PCR analysis without the need for further manipulations or optimisation of the PCR reaction mix, and it increased the reproducibility and specificity of the IgH PCR products.

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