Preselection of shotgun clones by oligonucleotide fingerprinting: an efficient and high throughput strategy to reduce redundancy in large-scale sequencing projects.
Large-scale genomic sequencing projects generally rely on random sequencing of shotgun clones, followed by different gap closing strategies. To reduce the overall effort and cost of those projects and to accelerate the sequencing throughput, we have developed an efficient, high throughput oligonucleotide fingerprinting protocol to select optimal shotgun clone sets prior to sequencing. Both computer simulations and experimental results, obtained from five PAC-derived shotgun libraries spanning 535 kb of the 17p11.2 region of the human genome, demonstrate that at least a 2-fold reduction in the number of sequence reads required to sequence an individual genomic clone (cosmid, PAC, etc.) can be achieved. Treatment of clone contigs with significant clone overlaps will allow an even greater reduction.
Radelof
U
U
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, 14195 Berlin, Germany. radelof@mpimp-berlin-dahlem.mpg.de
Hennig
S
S
Seranski
P
P
Steinfath
M
M
Ramser
J
J
Reinhardt
R
R
Poustka
A
A
Francis
F
F
Lehrach
H
H
D002886:Chromosomes, Human, Pair 17; D003001:Cloning, Molecular / Q000379:methods*; D003198:Computer Simulation; D016172:DNA Fingerprinting / Q000379:methods*; D015698:Genomic Library; D006801:Humans; D009841:Oligonucleotides / Q000235:genetics*; D010865:Pilot Projects; D017422:Sequence Analysis, DNA / Q000379:methods*
D009841:Oligonucleotides