B. Blomeke et al., SERUM, PLASMA AND PARAFFIN-EMBEDDED TISSUES AS SOURCES OF DNA FOR STUDYING CANCER SUSCEPTIBILITY GENES, Carcinogenesis, 18(6), 1997, pp. 1271-1275
The ability to isolate DNA from archived human serum, plasma and paraf
fin-embedded human tissues enhances opportunities to study breast, lun
g and other cancer risk factors, We report herein a simple and fast pr
otocol for the extraction of genomic DNA from these sources, Using a p
henol-based extraction method, the recovery for DNA is quantitative an
d reproducible, DNA yields in serum (250 mu l) were between 162 and 10
60 ng (It = 18 subjects), in plasma (250 mu l) were between 165 and 37
5 ng (it = 5 subjects) and in embedded tissues (5-mu m thick sections
for ethanol fixed, and between 5- and 20-mu m sections for formaldehyd
e fixation) were between 1 mu g and 11.7 mu g (it = 32 subjects), The
extraction method was combined with newly designed PCR-based assays fo
r cancer susceptibility marker genes such as CYP1A1 (exon 7), CYP2E1 (
Dra1, Rsa1), GSTM1 and NAT2 [NAT25A ((CT)-T-481), NAT2*6A (G(590)A),
NAT27A (G(857)A)I] Genotyping results from the serum and paraffin-emb
edded tissues compared favorably to results from archived freshly froz
en tissues, where concordance was 98% for serum, 100% for ethanol-fixe
d embedded tissues, and 97% for formaldehyde-fixed and paraffin-embedd
ed tissues, This facile method will allow for the use of archived tiss
ue samples of prospective cohort and other studies where intact DNA wa
s not previously available.