Laser microdissection for generation of a human chromosome region-specific library

被引:22
作者
He, W
Liu, YG
Smith, M
Berns, MW
机构
[1] UNIV CALIF IRVINE,BECKMAN LASER INST & MED CLIN,IRVINE,CA 92612
[2] UNIV CALIF IRVINE,DEPT PEDIAT,IRVINE,CA 92612
关键词
laser microdissection; region-specific DNA library; microcloning; laser trapping; chromosome targeting;
D O I
10.1017/S1431927697970033
中图分类号
T [工业技术];
学科分类号
08 [工学];
摘要
A human chromosome pq34 region-specific, microdissected library was constructed by using laser microdissection techniques. This library contains over 10,000 clones with an average insert size of 450 bp. It has greater coverage of the dissected chromosome region as compared with the needle-dissected chromosome 9q34 library. The laser microdissection technique provides more accurate chromosome targeting and easier operation than existing needle microdissection techniques. To simplify the procedure for chromosome microdissection and chromosome fragment collection, a trapping-cutting system was developed. This technique involves the use of two trapping beams which hold a single chromosome in suspension, and a third cutting beam, which dissects the immobilized chromosome. A collection chamber allowing for the fast collection of dissected chromosome fragments needs to be developed. However, DNA, can be cloned from trapped chromosome fragments with an insert size comparable to that of both needle-cut and laser-cut chromosomes.
引用
收藏
页码:47 / 52
页数:6
相关论文
共 8 条
[1]
IN VITRO PRODUCTION OF CHROMOSOMAL LESIONS WITH AN ARGON LASER MICROBEAM [J].
BERNS, MW ;
OLSON, RS ;
ROUNDS, DE .
NATURE, 1969, 221 (5175) :74-&
[2]
Buckle VJ, 1993, HUMAN GENETIC DISEAS, P59
[3]
JONSON DH, 1990, GENOMICS, V6, P243
[4]
CLONING DEFINED REGIONS OF THE HUMAN GENOME BY MICRODISSECTION OF BANDED CHROMOSOMES AND ENZYMATIC AMPLIFICATION [J].
LUDECKE, HJ ;
SENGER, G ;
CLAUSSEN, U ;
HORSTHEMKE, B .
NATURE, 1989, 338 (6213) :348-350
[5]
RAPID GENERATION OF REGION SPECIFIC PROBES BY CHROMOSOME MICRODISSECTION AND THEIR APPLICATION [J].
MELTZER, PS ;
GUAN, XY ;
BURGESS, A ;
TRENT, JM .
NATURE GENETICS, 1992, 1 (01) :24-28
[6]
MICRODISSECTION OF HUMAN-CHROMOSOMES BY A LASER MICROBEAM [J].
MONAJEMBASHI, S ;
CREMER, C ;
CREMER, T ;
WOLFRUM, J ;
GREULICH, KO .
EXPERIMENTAL CELL RESEARCH, 1986, 167 (01) :262-265
[7]
MICRO-DISSECTION AND CLONING OF DNA FROM A SPECIFIC REGION OF DROSOPHILA-MELANOGASTER POLYTENE CHROMOSOMES [J].
SCALENGHE, F ;
TURCO, E ;
EDSTROM, JE ;
PIRROTTA, V ;
MELLI, M .
CHROMOSOMA, 1981, 82 (02) :205-216
[8]
DEGENERATE OLIGONUCLEOTIDE-PRIMED PCR - GENERAL AMPLIFICATION OF TARGET DNA BY A SINGLE DEGENERATE PRIMER [J].
TELENIUS, H ;
CARTER, NP ;
BEBB, CE ;
NORDENSKJOLD, M ;
PONDER, BAJ ;
TUNNACLIFFE, A .
GENOMICS, 1992, 13 (03) :718-725