Direct STR amplification from whole blood and blood- or saliva-spotted FTA without DNA purification.
The DNA purification step has been thought to be essential for typing of STR DNA. However, this process is time-consuming, and there is a risk of unexpected cross-contamination during purification. We report a new method for direct short tandem repeat (STR) amplification using a newly developed direct PCR buffer, AnyDirect, which can amplify STR loci from whole blood and blood- or saliva-spotted FTA cards without DNA purification. The autosomal and Y chromosomal STR loci were analyzed for whole blood and blood or saliva spots of random individuals, followed by comparison of the results with those of corresponding purified DNA. The results from whole blood and blood spots showed perfect concordance with those from purified DNA without allele or locus drop-out. However, in the case of saliva spots, no amplification or locus drop-out was observed in some of the samples, which offers a topic for further study. Additionally, some commercial hot-start DNA polymerases other than AmpliTaq Gold DNA polymerase were also found to be compatible with this buffer system. Therefore, this direct PCR buffer was demonstrated to be useful for fast forensic DNA analysis or criminal DNA databases for which there is no need to store DNA samples.
Park
Su Jeong
SJ
DNA Analysis Lab, Forensic Science Division, Supreme Prosecutors' Office, Seocho-Gu, Seoul, Korea.
Kim
Jong Yeol
JY
Yang
Young Geun
YG
Lee
Seung Hwan
SH
D004247:DNA / Q000097:blood*; D016172:DNA Fingerprinting / Q000379:methods; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012463:Saliva / Q000378:metabolism*; D020080:Tandem Repeat Sequences
D004247:DNA