Laser pressure catapulting followed by B actin gene identification in Japanese quail macrochromosomes and microchromosomes using teflon‐coated coverslip slides

Abstract

Laser microdissection of individual mammalian chromosomes (> 2 microm) has been achieved though the use of a microscope slide coated with a polyethylene naphthalate (PEN) membrane. Although these slides have proved sufficient for larger chromosomes, they are insufficient for small chromosomes (< 1 microm). We have developed a new type of slide which allows laser microdissection of single Japanese quail microchromosomes (0.5 microm) and macrochromosomes (3-4 microm). To test the usefulness of these slides, a Japanese quail single nucleus, a macrochromosome, and a microchromosome were collected with Laser pressure catapulting, the B-actin gene was PCR amplified, and sequenced. The resulting PCR product was confirmed by nucleotide sequencing to be B-actin. These newly developed slides were shown to facilitate the laser microdissection of both Japanese quail macrochromosomes and microchromosomes.