[Laser chromosome microdissection and cloning of the genetic disease loci].
We have developed an argon ion laser chromosome microdissection technique in conjunction with a single unique primer polymerase chain reaction (SUP-PCR) to directly amplify microdissected chromosomes. The 22-mer primer used in PCR, although unique in sequence, randomly primed and amplified any target DNA. These methods were applied to both the terminal region of the human chromosome 4p (4p 16) and Xq (Xq26-q28), and two chromosome region-specific DNA libraries were constructed. The resulting libraries contained approximately 1000 nonoverlapping DNA sequences with an average size of 230-350 bp, at an average spacing of 10-65 Kbp along the chromosomes of origin. Our new method is a simple and general approach for constructing a chromosome region-specific DNA library from a single metaphase spread.
Hadano
S
S
Ikeda GenoSPHERE Project, ERATO, JRDC.
Ikeda
J E
JE
D001483:Base Sequence; D002894:Chromosomes, Human, Pair 4; D003001:Cloning, Molecular; D004247:DNA; D015698:Genomic Library; D006801:Humans; D007834:Lasers; D008846:Micromanipulation / Q000379:methods; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014960:X Chromosome
D004247:DNA