RESUMO
We used a novel DNA fingerprinting probe O-chi-1 (ref. 1) to detect differences in the hybridization pattern of brain tumor DNA and paired normal tissue of a given individual. Representatives of meningiomas (two), glioblastoma multeforme (three) and astrocytoma (one) were studied. Alterations, which included amplification as well as the loss of a normal band in tumor DNA, were observed in four of the six tumours. While the increased intensity of a band can be taken to imply increased copy number, the disappearance of bands could either be due to loss of DNA sequence or rearrangement resulting in different sized bands.
Assuntos
Astrocitoma/genética , Southern Blotting , Neoplasias Encefálicas/genética , Impressões Digitais de DNA/métodos , Sondas de DNA , DNA de Neoplasias/química , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Eletroforese em Gel de Ágar , Glioblastoma/genética , Humanos , Meningioma/genéticaRESUMO
Various storage treatments on human blood samples have been described with respect to DNA yield, quality and fingerprinting. Blood samples were stored at room temperature, 4 degrees C, -20 degrees C and -70 degrees C for different duration varying from 1, 2, 4 and 8 weeks with or without anticoagulant/preservative (EDTA or Heparin). DNA was isolated from these stored samples, quantitated by spectrophotometry and subjected to DNA fingerprinting using a human alphoid satellite DNA sequence (TRF 5.6) and a 33 mer oligonucleotide (O-chi-1) as probes. The polymorphic DNA bands were apparent between 2 to 9 kb size range and the fingerprints were individual-specific. Our results suggest that higher amount of genomic DNA can be recovered from blood samples stored at temperatures 4 degrees C or below in the presence of EDTA or heparin.