چكيده لاتين :
Background: Several methods have been developed for detection of sequence variation in genes and each has
its advantages and disadvantages. A disadvantage of them is that the simpler, cost-effective methods are
commonly perceived as being less sensitive in their detection of sequence variation, whereas those with
proven sensitivity have a requirement for complex or expensive laboratory equipment. In this context, we
undertook improvements to the conformation sensitive gel electrophoresis (CSGE) method which provides a
cost-effective approach to mutation detection and compared the results with scanning carried out using
denaturing high performance liquid chromatography (DHPLC) which utilises a dedicated analyser. Methods:
We designed PCR primers to amplify the seven protein-coding exons of the human SPP2 gene which encodes
secreted phosphoprotein 24 (spp24) such that the amplified products included the immediately-adjacent
intronic regions. Five improvements were made to the CSGE method that was used to the scan the PCRamplified
DNA. The scanning was then repeated using DHPLC and the results were compared. Results:
Using CSGE, a single nucleotide polymorphism was discovered in exon 2 and another in intron 2 of the gene.
Re-scanning of the same regions by DHPLC detected no additional sequence polymorphisms. Conclusion:
With modification of the original protocol, CSGE is capable of providing a simple and cost-effective approach
to the detection of DNA sequence polymorphisms that appears to be comparable in sensitivity to DHPLC.
Iran. Biomed. 1. 12 (2): 109-114, 2008
