1995
DOI: 10.1002/rcm.1290090905
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Rapid screening of genetic polymorphisms using buccal cell DNA with detection by matrix‐assisted laser desorption/ionization mass spectrometry

Abstract: A new approach is developed for the rapid and cost-effective detection of human genetic polymorphisms based on matrix-assisted laser description/ionization mass spectrometric (MALDI MS) detection using a nitrocellulose film substrate. This method employs polymerase chain reaction (PCR) amplification using DNA extracted from buccal cells as templates, followed by direct digestion with restriction enzymes and subsequent analysis by MALDI MS. The extraction of DNA from buccal cells provides a rapid and convenient… Show more

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Cited by 81 publications
(68 citation statements)
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“…The product is transferred onto the MALDI target without purification. This is in stark contrast to any other SNP genotyping method using MALDI as the detection system (7)(8)(9)(10)(11)(12)(13)(14)(15)(16). Of the starting PCR <0.5% are used for the MALDI sample preparation and in the MALDI analysis only a fraction of this is used.…”
Section: Discussionmentioning
confidence: 98%
See 1 more Smart Citation
“…The product is transferred onto the MALDI target without purification. This is in stark contrast to any other SNP genotyping method using MALDI as the detection system (7)(8)(9)(10)(11)(12)(13)(14)(15)(16). Of the starting PCR <0.5% are used for the MALDI sample preparation and in the MALDI analysis only a fraction of this is used.…”
Section: Discussionmentioning
confidence: 98%
“…MALDI has been applied to the analysis of DNA in variations that range from the analysis of PCR products (7,8), to approaches using sequencing (9), allele specific termination (10), single nucleotide primer extension reactions (11)(12)(13)(14) and hybridisation with PNAs (15,16). The major drawback of these approaches is that they heavily rely on stringent purification procedures prior to MALDI analysis that do not lend themselves to easy automation and contribute to the cost of a reaction.…”
Section: Introductionmentioning
confidence: 99%
“…MS directly assesses the nature of the PCR products, either as a whole or as fragmented oligonucleotides, whereas other technologies only indirectly measure PCR products, either through hybridization or by primer-extended minisequencing reactions, which use PCR products as templates. Genotyping by creating or abolishing recognition sites for restriction enzymes, similar to conventional restriction fragment length polymorphism analysis, has been used in combination with MALDI MS detection (28 ). Either naturally occurring restriction sites were used or base changes were incorporated in one of the PCR primers to create a recognition site with one of the alleles of the polymorphic site.…”
Section: Discussionmentioning
confidence: 99%
“…2 For nucleotide base substitutions, altered cleavage due to a change in the recognition sequence of a specific restriction enzyme has been utilized to detect genetic polymorphisms in carbonic anhydrase and CFTR genes. 3 The same strategy has been applied to the detection of another type of hexosaminidase A gene mutation. 2 However, direct identification of a molecular mass change due to substitution in a PCR product by MALDI-TOFMS has not been reported to date.…”
mentioning
confidence: 98%