Technical note: Efficiency of total demineralization and ion-exchange column for DNA extraction from bone.
We investigated whether a combination of recently introduced methods, total demineralization and ion-exchange columns, would increase DNA recovery from old bone. Ten bone samples taken after a burial period of approximately 60 years were used in this study. Bone powder was digested using total or incomplete demineralization. DNA was extracted by the standard organic method. The DNA extract was purified with ion-exchange columns or QIAquick spin columns. The efficiency of different DNA extraction methods was compared in terms of DNA concentration, inhibitors generated by real-time PCR, and conventional STR typing results. The mean DNA concentration using the total demineralization method is approximately 3 times higher than that using the incomplete demineralization method. For DNA purification, the method using QIAquick spin columns appeared to yield approximately double the DNA than the method using ion-exchange columns. Furthermore, 2 out of 10 samples showed higher levels of inhibition with C(T) values of IPC > or =30 cycles when using only ion-exchange columns. In STR results, total demineralization yielded more locus profiles by 4.2 loci than incomplete demineralization, and QIAquick spin columns also yielded more locus profiles by 3.5 loci than ion-exchange columns. Total demineralization of bone powder significantly increased DNA yield and improved STR typing results. However, the use of ion-exchange columns was not efficient when compared with the method using QIAquick spin columns. It is suggested that the combination of total demineralization and QIAquick spin columns lead to greatly improved STR typing results.
Seo
Seung Bum
SB
Department of Forensic Medicine, College of Medicine, Seoul National University, Seoul 110-799, South Korea.
Zhang
Aihua
A
Kim
Hye Yeon
HY
Yi
Jin A
JA
Lee
Hye Young
HY
Shin
Dong Hoon
DH
Lee
Soong Deok
SD
D018775:Bone Demineralization Technique; D001842:Bone and Bones / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D018732:Forensic Anthropology / Q000379:methods; D006801:Humans; D016133:Polymerase Chain Reaction
D004247:DNA