Analysis of methylation and distribution of CpG sequences on human active and inactive X chromosomes by in situ nick translation.
In situ nick translation of fixed mitotic chromosomes after HpaII or MspI digestion allows us to detect different DNA methylation levels along chromosomes. We used this technique to analyse the methylation levels of CCGG sites in the active and inactive X chromosomes of female human cells. In addition, we analysed the distribution of these sites with respect to the banding pattern. Our data show that the inactive X, as a whole, is more methylated than the active one and that CCGG sequences are preferentially located on R-positive bands.
Prantera
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Dipartimento di Genetica e Biologia Molecolare, Università La Sapienza, Roma, Italy.
Ferraro
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D001973:Bromodeoxyuridine / Q000378:metabolism; D002460:Cell Line; D002871:Chromosome Banding; D004247:DNA / Q000378:metabolism*; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000032:analysis / Q000378:metabolism*; D005260:Female; D006801:Humans; D008745:Methylation; D014960:X Chromosome / Q000378:metabolism*
D001973:Bromodeoxyuridine; D004247:DNA; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D015226:Dinucleoside Phosphates; C015772:CpG dinucleotide