Parallel DNA template preparation using a vacuum filtration sample transfer device.
Solid-phase techniques have facilitated the handling of biochemical analytes. This has stimulated the development of systems by which large sample panels can be analyzed with high levels of security and quality. We describe a sample transfer device based on the principle of vacuum filtration, which enables parallel handling of 96 samples of analytes bound to Sepharose beads. The tool was employed for strand separation of DNA samples, by attracting the beads to filter probes while passing them between the reagent solutions. The samples were analyzed using Pyrosequencing technology and proved to yield genotyping results of high quality. The presented sample preparation procedure provides an important link in the development of integrated systems for rapid genetic analysis at a low cost. In addition, the same filter could be reused extensively with very low risk for detectable cross-contamination between assays and without any reduction in processing capacity, thus further reducing the cost per analyzed sample.
Dunker
J
J
Pyrosequencing AB, Uppsala, Sweden. jenny.dunker@pyrosequencing.com
Larsson
U
U
Petersson
D
D
Forsell
J
J
Schiller
A L
AL
Alderborn
A
A
Berg
L M
LM
D004247:DNA / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D008846:Micromanipulation / Q000295:instrumentation* / Q000379:methods*; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods*; D014462:Ultrafiltration / Q000379:methods*; D014618:Vacuum
D004247:DNA