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.
Genome Sequencing Platform, Broad Institute of MIT and Harvard, 320 Charles St, Cambridge, MA 02141, USA.
Niall J
NJ
Lennon
Robert E
RE
Lintner
Scott
S
Anderson
Pablo
P
Alvarez
Andrew
A
Barry
William
W
Brockman
Riza
R
Daza
Rachel L
RL
Erlich
Georgia
G
Giannoukos
Lisa
L
Green
Andrew
A
Hollinger
Cindi A
CA
Hoover
David B
DB
Jaffe
Frank
F
Juhn
Danielle
D
McCarthy
Danielle
D
Perrin
Karen
K
Ponchner
Taryn L
TL
Powers
Kamran
K
Rizzolo
Dana
D
Robbins
Elizabeth
E
Ryan
Carsten
C
Russ
Todd
T
Sparrow
John
J
Stalker
Scott
S
Steelman
Michael
M
Weiand
Andrew
A
Zimmer
Matthew R
MR
Henn
Chad
C
Nusbaum
Robert
R
Nicol
D000465:Algorithms; D001330:Electronic Data Processing; D015723:Gene Library; D057166:High-Throughput Screening Assays; D006801:Humans; D008863:Microspheres; D017422:Sequence Analysis, DNA / Q000379:methods*