Development of a sensitive PCR to detect allele loss in a model hematopoietic neoplasm

J. Fairman, David Claxton, C. L. Willman, A. B. Deisseroth, L. Nagarajan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Loss or gain of an entire chromosome and interstitial deletions or amplification are hallmarks of several hematopoietic neoplasms. These chromosomal anomalies can be identified by conventional cytogenetic analysis of bone marrow aspirates. We have developed a PCR-based assay to detect loss of chromosome 5q31 loci, in the model system of myeloid disorders with the 5q- chromosome (interstitial deletion of 5q), by taking advantage of a highly polymorphic dinucleotide repeat within the interleukin-9 (IL9) gene on 5q31. In a given sample, quantitation of amplification of individual alleles in a Phosphorlmager allowed the representation of alleles to be expressed as a ratio of the larger to the smaller allele. Comparison of these ratios in paired DNA samples from Ficoll buoyant and pelletted fractions provides evidence for allele loss. Results presented here demonstrate that this technique of comparison of ratios of isotope incorporation could be expanded to investigate any deletion or numerical abnormality in hematopoietic tumors.

Original languageEnglish (US)
Pages (from-to)6-12
Number of pages7
JournalPCR Methods and Applications
Volume4
Issue number1
StatePublished - Dec 1 1994

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Hematologic Neoplasms
Alleles
Polymerase Chain Reaction
Chromosome Deletion
Interleukin-9
Dinucleotide Repeats
Ficoll
Cytogenetic Analysis
Isotopes
Chromosomes
Bone Marrow
DNA
Genes
Neoplasms

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Cite this

Fairman, J., Claxton, D., Willman, C. L., Deisseroth, A. B., & Nagarajan, L. (1994). Development of a sensitive PCR to detect allele loss in a model hematopoietic neoplasm. PCR Methods and Applications, 4(1), 6-12.
Fairman, J. ; Claxton, David ; Willman, C. L. ; Deisseroth, A. B. ; Nagarajan, L. / Development of a sensitive PCR to detect allele loss in a model hematopoietic neoplasm. In: PCR Methods and Applications. 1994 ; Vol. 4, No. 1. pp. 6-12.
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Fairman, J, Claxton, D, Willman, CL, Deisseroth, AB & Nagarajan, L 1994, 'Development of a sensitive PCR to detect allele loss in a model hematopoietic neoplasm', PCR Methods and Applications, vol. 4, no. 1, pp. 6-12.

Development of a sensitive PCR to detect allele loss in a model hematopoietic neoplasm. / Fairman, J.; Claxton, David; Willman, C. L.; Deisseroth, A. B.; Nagarajan, L.

In: PCR Methods and Applications, Vol. 4, No. 1, 01.12.1994, p. 6-12.

Research output: Contribution to journalArticle

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N2 - Loss or gain of an entire chromosome and interstitial deletions or amplification are hallmarks of several hematopoietic neoplasms. These chromosomal anomalies can be identified by conventional cytogenetic analysis of bone marrow aspirates. We have developed a PCR-based assay to detect loss of chromosome 5q31 loci, in the model system of myeloid disorders with the 5q- chromosome (interstitial deletion of 5q), by taking advantage of a highly polymorphic dinucleotide repeat within the interleukin-9 (IL9) gene on 5q31. In a given sample, quantitation of amplification of individual alleles in a Phosphorlmager allowed the representation of alleles to be expressed as a ratio of the larger to the smaller allele. Comparison of these ratios in paired DNA samples from Ficoll buoyant and pelletted fractions provides evidence for allele loss. Results presented here demonstrate that this technique of comparison of ratios of isotope incorporation could be expanded to investigate any deletion or numerical abnormality in hematopoietic tumors.

AB - Loss or gain of an entire chromosome and interstitial deletions or amplification are hallmarks of several hematopoietic neoplasms. These chromosomal anomalies can be identified by conventional cytogenetic analysis of bone marrow aspirates. We have developed a PCR-based assay to detect loss of chromosome 5q31 loci, in the model system of myeloid disorders with the 5q- chromosome (interstitial deletion of 5q), by taking advantage of a highly polymorphic dinucleotide repeat within the interleukin-9 (IL9) gene on 5q31. In a given sample, quantitation of amplification of individual alleles in a Phosphorlmager allowed the representation of alleles to be expressed as a ratio of the larger to the smaller allele. Comparison of these ratios in paired DNA samples from Ficoll buoyant and pelletted fractions provides evidence for allele loss. Results presented here demonstrate that this technique of comparison of ratios of isotope incorporation could be expanded to investigate any deletion or numerical abnormality in hematopoietic tumors.

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