Fig. 4.
Fig. 4. Analysis of X chromosome inactivation. / DNA was isolated from PBMCs and after restriction digestion with eitherRsaI and HpaII or RsaI alone, was amplified using primers specific for the sequences flanking methylation sensitive restriction sites in exon 1 of the androgen receptor gene. Amplified products were electrophoresed on nondenaturing gel and detected by silver stain. In the RsaI digest both active (unmethylated) and inactive (methylated) alleles are amplified, whereas after HpaII digest only inactive allele is amplified. Random X inactivation is identified by presence of 2 bands afterHpaII digestion; this pattern was found for the patient. In contrast, the patient's mother has only one band, indicating nonrandom inactivation. As anticipated, only one band was found inRsaI digests of the father's DNA.

Analysis of X chromosome inactivation.

DNA was isolated from PBMCs and after restriction digestion with eitherRsaI and HpaII or RsaI alone, was amplified using primers specific for the sequences flanking methylation sensitive restriction sites in exon 1 of the androgen receptor gene. Amplified products were electrophoresed on nondenaturing gel and detected by silver stain. In the RsaI digest both active (unmethylated) and inactive (methylated) alleles are amplified, whereas after HpaII digest only inactive allele is amplified. Random X inactivation is identified by presence of 2 bands afterHpaII digestion; this pattern was found for the patient. In contrast, the patient's mother has only one band, indicating nonrandom inactivation. As anticipated, only one band was found inRsaI digests of the father's DNA.

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