TY - JOUR
T1 - Identification of human DNA sequences complementary to chromosomal RNA with an improved RNA/DNA gradient hybridization technique
AU - Tsai, Michael Y.
AU - Yunis, Jorge J.
N1 - Funding Information:
This research was supported by Grant No. HD-01962 from the National Institutes of Health.
PY - 1977/4/4
Y1 - 1977/4/4
N2 - Native DNA components in the human genome were identified with an improved RNA gradient hybridization technique. 3H-Labelled complementary RNA transcribed from total human DNA was hybridized to homologous DNA which had been mildly sheared to generate single-stranded ends. Unhybridized RNA was eliminated from the reaction mixture by column chromatography before the hybrids were subjected to CsCl gradient centrifugation. Nine radioactive peaks representing RNA hybrids were reproducibly found at buoyant densities of 1.687, 1.693, 1.696, 1.699, 1.702, 1.705, 1.708, 1.711 and 1.715 g/cm3. The technique was also utilized to hybridize 125I-labelled human chromosomal RNA to DNA. Three distinct radioactive peaks were found at the heavy buoyant densities of 1.702, 1.705, and 1.708 g/cm3, suggesting a differential enrichment of DNA components with sequences complementary to chromosomal RNA.
AB - Native DNA components in the human genome were identified with an improved RNA gradient hybridization technique. 3H-Labelled complementary RNA transcribed from total human DNA was hybridized to homologous DNA which had been mildly sheared to generate single-stranded ends. Unhybridized RNA was eliminated from the reaction mixture by column chromatography before the hybrids were subjected to CsCl gradient centrifugation. Nine radioactive peaks representing RNA hybrids were reproducibly found at buoyant densities of 1.687, 1.693, 1.696, 1.699, 1.702, 1.705, 1.708, 1.711 and 1.715 g/cm3. The technique was also utilized to hybridize 125I-labelled human chromosomal RNA to DNA. Three distinct radioactive peaks were found at the heavy buoyant densities of 1.702, 1.705, and 1.708 g/cm3, suggesting a differential enrichment of DNA components with sequences complementary to chromosomal RNA.
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U2 - 10.1016/0005-2787(77)90057-0
DO - 10.1016/0005-2787(77)90057-0
M3 - Article
C2 - 856272
AN - SCOPUS:0017363443
SN - 0005-2787
VL - 475
SP - 417
EP - 423
JO - BBA Section Nucleic Acids And Protein Synthesis
JF - BBA Section Nucleic Acids And Protein Synthesis
IS - 3
ER -