A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454.
We present an automated, high throughput library construction process for 454 technology. Sample handling errors and cross-contamination are minimized via end-to-end barcoding of plasticware, along with molecular DNA barcoding of constructs. Automation-friendly magnetic bead-based size selection and cleanup steps have been devised, eliminating major bottlenecks and significant sources of error. Using this methodology, one technician can create 96 sequence-ready 454 libraries in 2 days, a dramatic improvement over the standard method.
Lennon
Niall J
NJ
Genome Sequencing Platform, Broad Institute of MIT and Harvard, 320 Charles St, Cambridge, MA 02141, USA.
Lintner
Robert E
RE
Anderson
Scott
S
Alvarez
Pablo
P
Barry
Andrew
A
Brockman
William
W
Daza
Riza
R
Erlich
Rachel L
RL
Giannoukos
Georgia
G
Green
Lisa
L
Hollinger
Andrew
A
Hoover
Cindi A
CA
Jaffe
David B
DB
Juhn
Frank
F
McCarthy
Danielle
D
Perrin
Danielle
D
Ponchner
Karen
K
Powers
Taryn L
TL
Rizzolo
Kamran
K
Robbins
Dana
D
Ryan
Elizabeth
E
Russ
Carsten
C
Sparrow
Todd
T
Stalker
John
J
Steelman
Scott
S
Weiand
Michael
M
Zimmer
Andrew
A
Henn
Matthew R
MR
Nusbaum
Chad
C
Nicol
Robert
R
D000465:Algorithms; D001330:Electronic Data Processing; D015723:Gene Library; D057166:High-Throughput Screening Assays; D006801:Humans; D008863:Microspheres; D017422:Sequence Analysis, DNA / Q000379:methods*