Cytogenetic analysis by chromosome painting using dop-pcr amplified flow-sorted chromosomes
✍ Scribed by HÅKan Telenius; Bruce A. J. Ponder; Alan Tunnacliffe; Adèle H. Pelmear; Nigel P. Carter; Malcolm A. Ferguson-Smith; Annemarie Behmel; Magnus Nordenskjöld; Roswitha Pfragner
- Publisher
- John Wiley and Sons
- Year
- 1992
- Tongue
- English
- Weight
- 670 KB
- Volume
- 4
- Category
- Article
- ISSN
- 1045-2257
No coin nor oath required. For personal study only.
✦ Synopsis
A novel polymerase chain reaction (PCR) technique has been combined with chromosome flow sorting to characterise two lymphoblastoid cell lines and one medullary thyroid carcinoma cell line carrying translocations close t o the locus for multiple endocrine neoplasia type 2A (MEN 2A). Five hundred copies of the derivative chromosome(s) were flow sorted from each cell line and amplified by degenerate oligonucleotide-primed-polymerase chain reaction (DOP-PCR). This generated pools of DNA sequences corresponding t o the abnormal chromosomes, which were then used as probes in fluorescence in situ hybridisation (FISH) experiments on normal metaphase cells. The resultant chromosome paints revealed the portions of the normal chromosomes related t o those involved in the translocations. By this technique, translocation breakpoints in bands p IS,