Skip to main content
NCBI home page
Molecular Pathology : MP logoLink to Molecular Pathology : MP
. 1997 Jun;50(3):164–166. doi: 10.1136/mp.50.3.164

Modification of IgH PCR clonal analysis by the addition of sucrose and cresol red directly to PCR reaction mixes.

E Hodges 1, S M Boddy 1, S Thomas 1, J L Smith 1
PMCID: PMC379613  PMID: 9292153

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.

Full text

PDF
164

Images in this article

165Image
on p.165

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bateman A. C., Sage D. A., Al-Talib R. K., Theaker J. M., Jones D. B., Howell W. M. Investigation of specimen mislabelling in paraffin-embedded tissue using a rapid, allele-specific, PCR-based HLA class II typing method. Histopathology. 1996 Feb;28(2):169–174. doi: 10.1046/j.1365-2559.1996.277323.x. [DOI] [PubMed] [Google Scholar]
  2. Diss T. C., Pan L., Peng H., Wotherspoon A. C., Isaacson P. G. Sources of DNA for detecting B cell monoclonality using PCR. J Clin Pathol. 1994 Jun;47(6):493–496. doi: 10.1136/jcp.47.6.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Greer C. E., Peterson S. L., Kiviat N. B., Manos M. M. PCR amplification from paraffin-embedded tissues. Effects of fixative and fixation time. Am J Clin Pathol. 1991 Feb;95(2):117–124. doi: 10.1093/ajcp/95.2.117. [DOI] [PubMed] [Google Scholar]
  4. Hodges E., Swindell A. M., Quin C. T., Lane A. C., Jones D. B., Sweetenham J., Smith J. L. Expanded TcR V beta 5.1 family in a diffuse high-grade B cell immunoblastic lymphoma. Leukemia. 1995 Jun;9(6):1108–1112. [PubMed] [Google Scholar]
  5. Hoppe B. L., Conti-Tronconi B. M., Horton R. M. Gel-loading dyes compatible with PCR. Biotechniques. 1992 May;12(5):679–680. [PubMed] [Google Scholar]
  6. Jackson D. P., Lewis F. A., Taylor G. R., Boylston A. W., Quirke P. Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction. J Clin Pathol. 1990 Jun;43(6):499–504. doi: 10.1136/jcp.43.6.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Potter M. N., Steward C. G., Maitland N. J., Oakhill A. Detection of clonality in childhood B-lineage acute lymphoblastic leukaemia by the polymerase chain reaction. Leukemia. 1992 Apr;6(4):289–294. [PubMed] [Google Scholar]

Articles from Molecular Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES