Sökning: onr:"swepub:oai:DiVA.org:uu-169821" >
Preimplantation dia...
Preimplantation diagnosis by whole-genome amplification, PCR amplification, and solid-phase minisequencing of blastomere DNA
-
Paunio, T (författare)
-
Reima, I (författare)
-
Syvänen, Ann-Christine (författare)
- 1996
- 1996
- Engelska.
-
Ingår i: Clinical Chemistry. - 0009-9147 .- 1530-8561. ; 42:9, s. 1382-1390
- Relaterad länk:
-
https://urn.kb.se/re...
Abstract
Ämnesord
Stäng
- We have developed a new method for preimplantation diagnosis of inherited diseases. Our procedure for the identification of point mutations in single cells combines whole-genome amplification using 15-mer random primers (primer extension preamplification, PEP) with a single locus-specific PCR amplification, followed by detection of the mutation by solid-phase minisequencing. The procedure was evaluated by detecting three disease-causing mutations and seven polymorphic nucleotides located on different human chromosomes from single granuloma and blastomere cells. The correct genotype of the cell was identified at 96% of the nucleotide positions analyzed, showing that a representative part of the genome is amplified during PEP. We estimate that PEP yielded at least 1000 copies of the genome. The quantitative nature of the solid-phase minisequencing method allowed us to notice that preferential amplification of one allele occurs at heterozygous loci during PEP, which is a potential problem in preimplantation diagnosis.
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas