Optimization of an automated DNA purification protocol for neonatal screening.
BACKGROUND
Collection of blood from newborns is a standard clinical procedure used for genetic screening. Typically, blood from a heel prick is absorbed onto standard collection paper and dried before analysis of metabolites, proteins, hormones, and more recently DNA.


OBJECTIVE
To evaluate strategies to purify DNA for use with automated workstations.


METHODS
Two factors were used to evaluate several DNA purification protocols: residual heme contamination and amplification yield. The protocol that produced DNA with the lowest heme content and the highest amplification yield was selected. In combination with those two performance factors, the protocol with the fewest number of steps was chosen to reduce reagent use and processing time.


METHODS
Industrial research and development laboratory.


RESULTS
Robust amplification of DNA isolated from dried blood spots was demonstrated using both fluorescence and agarose gel-based detection methods. In addition, the samples had consistent DNA volumes and had no detectable cross-contamination. Suggested instrument settings, equipment, and supplies were included for automated processing of DNA from dried blood spots.


CONCLUSIONS
A 4-step DNA processing protocol was developed for dried blood spots. The protocol could be performed in either a manual or automated format, making it possible to process hundreds of samples in 1 day.
Heath
E M
EM
Gentra Systems, Inc, Minneapolis, MN 55441, USA. eheath@gentra.com
O'Brien
D P
DP
Banas
R
R
Naylor
E W
EW
Dobrowolski
S
S
D001774:Blood Chemical Analysis / Q000379:methods; D004247:DNA / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D003936:Diagnosis, Computer-Assisted / Q000592:standards*; D005069:Evaluation Studies as Topic; D005820:Genetic Testing / Q000379:methods*; D006418:Heme / Q000032:analysis; D006801:Humans; D015997:Neonatal Screening / Q000379:methods*; D016133:Polymerase Chain Reaction
D006418:Heme; D004247:DNA