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Use of a biotinylated DNA probe specific for the human Y chromosome for rapid antenatal sex determination
Author(s) -
Boehm Thomas L. J.,
Drahovsky Dusan
Publication year - 1986
Publication title -
clinical genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 102
eISSN - 1399-0004
pISSN - 0009-9163
DOI - 10.1111/j.1399-0004.1986.tb01919.x
Subject(s) - clone (java method) , microbiology and biotechnology , biotinylation , hybridization probe , biology , dna , amniotic fluid , taqi , molecular probe , y chromosome , genetics , fetus , restriction fragment length polymorphism , polymerase chain reaction , gene , pregnancy
We have cloned a male‐specific 3.4 kb human DNA sequence which showed only little cross‐hybridisation to autosomal sequences. To further enhance the specificity of the probe, we subcloned an internal Taql‐fragment resulting in clone pH343T33. This clone was used to determine the presence of Y‐chromosomal sequences in DNA extracted from amniotic cells and from chorionic villi by Southern and dot hybridisation assays, respectively. Using this clone, we correctly predicted fetal sex in all of 148 cases analysed. To facilitate the use of this clone in clinical practice, we simplified the dot hybridisation procedure so that it can be performed in less than 48 h. The procedure with 32 P‐labeled DNA probes requires less than 0.5 mL of amniotic fluid; when biotinylated DNA probes are used, 3–5 mL of amniotic fluid usually suffice. We have used this probe in genetic counseling of families at risk for X‐linked disorders.