Critical factors in the performance and cost of two-dimensional gene scanning

RB1 as a model

R. K. Dhanda, N. J. Van Orsouw, Sigalas, C. Eng, Jan Vijg

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Two dimensional (2-D) gene scanning (TDGS) is a method for mutation detection based on the electrophoretic separation of PCR-amplified DNA fragments according to size and base pair sequence. The use of denaturing gradient gel electrophoresis (DGGE) as the second separation step provides virtually 100% sensitivity, while the 2-D format allows the inspection of multiple gene fragments simultaneously. Analysis of many exons in parallel is greatly facilitated by extensive PCR multiplexing based on preamplification by long-distance PCR. Recently, TDGS has been applied to detect mutations in the retinoblastoma tumor suppressor gene RB1. Using RB1 as a model, we have now analyzed each step of the protocol, presenting overall improvements and a detailed cost analysis, where the total cost of the assay is found to be about $40 (US). An overall picture of TDGS cost-performance, as compared to direct sequencing, is provided as a function of the number of target fragments.

Original languageEnglish (US)
Pages (from-to)664-675
Number of pages12
JournalBioTechniques
Volume25
Issue number4
StatePublished - 1998
Externally publishedYes

Fingerprint

Genes
Scanning
Costs and Cost Analysis
Polymerase Chain Reaction
Costs
Denaturing Gradient Gel Electrophoresis
Mutation
Retinoblastoma
Tumor Suppressor Genes
Electrophoresis
Multiplexing
Base Pairing
Tumors
Exons
Assays
Inspection
Gels
DNA

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Dhanda, R. K., Van Orsouw, N. J., Sigalas, Eng, C., & Vijg, J. (1998). Critical factors in the performance and cost of two-dimensional gene scanning: RB1 as a model. BioTechniques, 25(4), 664-675.

Critical factors in the performance and cost of two-dimensional gene scanning : RB1 as a model. / Dhanda, R. K.; Van Orsouw, N. J.; Sigalas; Eng, C.; Vijg, Jan.

In: BioTechniques, Vol. 25, No. 4, 1998, p. 664-675.

Research output: Contribution to journalArticle

Dhanda, RK, Van Orsouw, NJ, Sigalas, Eng, C & Vijg, J 1998, 'Critical factors in the performance and cost of two-dimensional gene scanning: RB1 as a model', BioTechniques, vol. 25, no. 4, pp. 664-675.
Dhanda, R. K. ; Van Orsouw, N. J. ; Sigalas ; Eng, C. ; Vijg, Jan. / Critical factors in the performance and cost of two-dimensional gene scanning : RB1 as a model. In: BioTechniques. 1998 ; Vol. 25, No. 4. pp. 664-675.
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