Abstract
A transmission electron microscope method for gene mapping by in situ hybridization to Drosophila polytene chromosomes has been developed. As electron-opaque labels, we use colloidal gold spheres having a diameter of 25 nm. The spheres are coated with a layer of protein to which Escherichia coli single-stranded DNA is photochemically crosslinked. Poly(dT) tails are added to the 3' OH ends of these DNA strands, and poly(dA) tails are added to the 3' OH ends of a fragmented cloned Drosophila DNA. These probe-dA strands are hybridized in situ to polytene chromosome squashes. Gold spheres are linked to the hybridized probe-dA strands by A.T base pairing. The sphere positions relative to the chromosome bands can be observed by transmission electron microscopy. The method shows low background and high resolution.
| Original language | English |
|---|---|
| Pages (from-to) | 7059-7063 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 78 |
| Issue number | 11 II |
| DOIs | |
| Publication status | Published - 1981 |
| Externally published | Yes |
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