Microdissection of human high resolution banded chromosome, polymerase chain reaction and microcloning.
A simple technique for microdissection of specific region of human high resolution banded chromosome, followed by polymerase chain reaction (PCR), and microcloning was successfully used to microdissect 4 chromosomal pieces from the distal one third from band 11.2 to the terminal of the short arm of Y chromosome where the testis determining factor is located; 3.6 x 10(4) clones were obtained after 30 cycles of PCR. We analysed 41 clones with insert. The size of insert ranges from 140 to 350 bp (average 250 bp). A Southern blot analysis was done for one of them, and a 2.5 kb Hind III fragment was detected.
Deng
H X
HX
National Laboratory of Medical Genetics, Hunan Medical University, Changsha.
He
X X
XX
Li
L Y
LY
Xia
J H
JH
Niikawa
N
N
D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004259:DNA-Directed DNA Polymerase; D004210:Dissection; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction; D019914:Taq Polymerase; D014998:Y Chromosome
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase