An efficient genomic DNA extraction from whole blood using Nextractor.
BACKGROUND
We evaluated the performance of the Nextractor NX-48 nucleic acid extractor system for the extraction of genomic DNA from whole blood samples.


METHODS
We compared the performance of the Nextractor to that of the QIAamp DNA Blood Mini Kit and the Maxwell system, using five whole blood samples. Extraction efficiencies were compared based on the total amount of extracted DNA adjusted by input blood volume, and the purity was compared. Polymerase chain reaction analyses were performed using ACTB as a target. The real-time PCR assay was carried out for housekeeping gene GAPDH. Total elapsed time for DNA extraction was compared.


RESULTS
Extraction efficiencies for the QIAamp, Maxwell, and Nextractor were 25.4±3.8ng/μL, 9.2±0.6ng/μL, and 31.0±5.6ng/μL, respectively. No significant differences in purity were observed among three methods. DNA extracted using the ACTB was successfully amplified in all three methods. There were no obvious differences in Ct values for GAPDH real-time PCR. Total elapsed time for DNA extraction was about 50min for the QIAamp, 40min for the Maxwell, and 20min for the Nextractor.


CONCLUSIONS
As the Nextractor is faster and requires less hands-on time than manual procedures, it may be useful for molecular diagnostic testing in clinical laboratories.
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea.
Tae-Dong
TD
Jeong
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea.
Young-Uk
YU
Cho
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea. Electronic address: wlee1@amc.seoul.kr.
Woochang
W
Lee
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea.
Sail
S
Chun
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea.
Won-Ki
WK
Min
D005591:Chemical Fractionation / Q000379:methods*; D004247:DNA / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification*; D015894:Genome, Human; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction
D004247:DNA
DNA extraction; Nextractor; Performance