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5 results on '"Bransfield, K."'

1. Real-time PCR based on SYBR-Green I fluorescence: An alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions

Author

Ponchel, F., Toomes, C., Bransfield, K., Leong, F.T., Douglas, S.H., Field, S.L., Bell, S.M., Combaret, V., Puisieux, A., Mighell, A.J., Robinson, P.A., Inglehearn, C.F., Isaacs, J.D., and Markham, A.F.

Abstract

BACKGROUND:\ud \ud Real-time PCR is increasingly being adopted for RNA quantification and genetic analysis. At present the most popular real-time PCR assay is based on the hybridisation of a dual-labelled probe to the PCR product, and the development of a signal by loss of fluorescence quenching as PCR degrades the probe. Though this so-called 'TaqMan' approach has proved easy to optimise in practice, the dual-labelled probes are relatively expensive.\ud \ud RESULTS:\ud \ud We have designed a new assay based on SYBR-Green I binding that is quick, reliable, easily optimised and compares well with the published assay. Here we demonstrate its general applicability by measuring copy number in three different genetic contexts; the quantification of a gene rearrangement (T-cell receptor excision circles (TREC) in peripheral blood mononuclear cells); the detection and quantification of GLI, MYC-C and MYC-N gene amplification in cell lines and cancer biopsies; and detection of deletions in the OPA1 gene in dominant optic atrophy.\ud \ud CONCLUSION:\ud \ud Our assay has important clinical applications, providing accurate diagnostic results in less time, from less biopsy material and at less cost than assays currently employed such as FISH or Southern blotting.\ud

Published
2003

2. Assignment<FOOTREF>[sup 1] </FOOTREF> of GALGT encoding β-1,4N-acetylgalactosaminyl-transferase (GalNAc-T) and KIF5A encoding neuronal kinesin (D12S1889) to human chromosome band 12q13 by assignment to ICI YAC 26EG10 and in situ hybridization.

Author

Hamlin, P. J., Jones, P. F., Leek, J. P., Bransfield, K., Lench, N. J., Aldersley, M. A., Howdle, P. D., Markham, A. F., and Robinson, P. A.

Subjects
HUMAN gene mapping, KINESIN, IN situ hybridization, EXONS (Genetics), MICROTUBULES, ACYLTRANSFERASES, HUMAN genetics, HUMAN chromosomes
Abstract

This article describes a cytogenetic experiment describing GALGT encoding beta-1,4N-acetylgalactosaminyl-transferase (GalNAc-T) and KIF5A encoding neuronal kinesin (D12S1889) to human chromosome band 12q13 by assignment to ICIYAC 26EG10 and in situ hybridization. The genes, 3- 1,4 N-acetylgalactosaminyl-transferase (GALGT) and human neuronal kinesin (KIF5A, Dl 2S 1889) were mapped. More specifically, GALGT and KIF5A map to the same 200-kb yeast artificial chromosome as GLI and DDIT3. Beta-1,4 N-acetylgalactosaminyltransferase (GM2/GD2 synthase) is an enzyme involved in the synthesis of complex gangliosides which have been suggested to have a role in neuronal functioning.

Published
1998
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3. Rubinstein-Taybi Syndrome: Dosage Analysis Using MLPA.

Author

Cook, Ian, Bransfield, K. L., Bennett, C. P., Cockburn, D. J., J. Williams, Taylor, G. R., and Delaney, C. A.

Subjects
INTELLECTUAL disabilities
Abstract

Presents an abstract of the article "Rubinstein-Taybi Syndrome: Dosage Analysis Using MLPA," by Ian Cook, K.L. Bransfield, C.P. Bennett, D.J. Cockburn, J. Williams, G.R. Taylor, and C.A. Delaney.

Published
2005

4. Real-time PCR based on SYBR-Green I fluorescence: an alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions.

Author

Ponchel F, Toomes C, Bransfield K, Leong FT, Douglas SH, Field SL, Bell SM, Combaret V, Puisieux A, Mighell AJ, Robinson PA, Inglehearn CF, Isaacs JD, and Markham AF

Subjects
Benzothiazoles, Cell Line, Tumor, DNA chemistry, Diamines, Female, Fluorescence, GTP Phosphohydrolases genetics, Gene Amplification genetics, Gene Deletion, Gene Rearrangement genetics, Humans, Male, Oncogene Proteins genetics, Optic Atrophy genetics, Quinolines, Receptors, Antigen, T-Cell genetics, Reproducibility of Results, Sensitivity and Specificity, Trans-Activators, Transcription Factors genetics, Zinc Finger Protein GLI1, DNA genetics, Organic Chemicals chemistry, Polymerase Chain Reaction methods
Abstract

Background: Real-time PCR is increasingly being adopted for RNA quantification and genetic analysis. At present the most popular real-time PCR assay is based on the hybridisation of a dual-labelled probe to the PCR product, and the development of a signal by loss of fluorescence quenching as PCR degrades the probe. Though this so-called 'TaqMan' approach has proved easy to optimise in practice, the dual-labelled probes are relatively expensive., Results: We have designed a new assay based on SYBR-Green I binding that is quick, reliable, easily optimised and compares well with the published assay. Here we demonstrate its general applicability by measuring copy number in three different genetic contexts; the quantification of a gene rearrangement (T-cell receptor excision circles (TREC) in peripheral blood mononuclear cells); the detection and quantification of GLI, MYC-C and MYC-N gene amplification in cell lines and cancer biopsies; and detection of deletions in the OPA1 gene in dominant optic atrophy., Conclusion: Our assay has important clinical applications, providing accurate diagnostic results in less time, from less biopsy material and at less cost than assays currently employed such as FISH or Southern blotting.

Published
2003
Full Text
View/download PDF

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