Use of a polymorphic dinucleotide repeat sequence to detect non-blastomeric contamination of the polymerase chain reaction in biopsy samples for preimplantation diagnosis.
Using the polymerase chain reaction (PCR), amplification of two different target DNA sequences has been achieved with high frequency using single human blastomeres as template for the duplex reaction. One sequence is located within the beta-globin gene and contains the sickle cell locus, the other is a polymorphic dinucleotide repeat, which, as well as acting as a positive control for amplification, was used to check the origin of the amplified DNA. A comparison of the sequences amplified from the blastomere with sequences amplified from parental samples confirmed that amplification of blastomeric sequences, but not extraneous contaminating DNA, had taken place in most cases. The efficacy of this system for detecting extraneous DNA was checked by deliberately contaminating single blastomeres with foreign cells. The presence of contamination was detected by the amplification of sequences not present in blastomeric DNA and which therefore must have been amplified from extraneous contaminating DNA.
Pickering
S J
SJ
Department of Anatomy, University of Cambridge, UK.
McConnell
J M
JM
Johnson
M H
MH
Braude
P R
PR
D000225:Adenine; D000755:Anemia, Sickle Cell / Q000235:genetics; D001483:Base Sequence; D001757:Blastomeres / Q000737:chemistry*; D003596:Cytosine; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D047108:Embryonic Development; D005260:Female; D005914:Globins / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000706:statistics & numerical data; D011786:Quality Control; D012091:Repetitive Sequences, Nucleic Acid
D003596:Cytosine; D005914:Globins; D004247:DNA; D000225:Adenine