Genomics 1992 Aug;13(4):1322-4
Section of Hematology/Oncology, University of Chicago, Illinois
We have developed a simple, efficient method by which
microdissected material can be amplified directly in the
collection container in a few hours. The procedure involves two
initial rounds of DNA synthesis with T7 DNA polymerase, using a
primer that contains a random pentanucleotide sequence at its 3'
end and a defined sequence at its 5' end, followed by PCR
amplification with the defined sequence as the primer. The
resulting products can be biotinylated and used for fluorescence
in situ hybridization (FISH) to confirm their chromosomal
location. As few as 17 dissected chromosomal regions provide
sufficient material for a specific FISH signal on the appropriate
band of metaphase chromosomes. We have obtained a chromosome
6q25-qter-specific painting probe in this way.
PMID: 1505965, UI: 92372035