Laser pressure catapulting followed by B actin gene identification in Japanese quail macrochromosomes and microchromosomes using teflon-coated coverslip slides.
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.
McNally
L R
LR
Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, 70803, USA.
Henk
W G
WG
Cooper
R K
RK
D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002458:Cell Fractionation / Q000379:methods*; D002875:Chromosomes; D016384:Consensus Sequence; D003370:Coturnix; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D008969:Molecular Sequence Data; D011138:Polytetrafluoroethylene; D012689:Sequence Homology, Nucleic Acid; D013499:Surface Properties
D000199:Actins; D011138:Polytetrafluoroethylene