TY - JOUR
T1 - Evaluating the performance of whole genome amplification for use in low template DNA typing
AU - Lee, James
AU - Tsai, Li-Chin
AU - Lai, Pi-Yi
AU - Lee, Cheng-Chang
AU - Lin, Chun-Yen
AU - Huang, Tsun-Ying
AU - Linacre, Adrian
AU - Hsieh, Hsing-Mei
PY - 2012/10
Y1 - 2012/10
N2 - We report on the performance of two whole genome amplification methods, GenomiPhiTM amplification and modifiedimproved primer extension preamplification (mIPEP), when analysing low template DNA samples. Template as low as 10 pg treated with mIPEP generated more than 1 ng of DNA that could be used in STR typing. Initial templates of 100-10 pg, when treated with mIPEP, generated an increase in alleles compared with control samples. Partial profiles using the AmpF'STRw IdentifilerTM Kit were produced from this suboptimal DNA template, with 70% of the possible alleles (21.7+2.1 in 32 alleles) recorded, using the mIPEP amplified products with an initial template of 100 pg. Allelic imbalance decreased with samples treated with whole genome amplification method (WGA) compared with those without this initial treatment. Further methods for improvement were also analysed including altering the condition of electrokinetic injection, and the successful DNA typing rate was increased to about 80%. This report illustrates the potential use and limitations of WGA for low template samples.
AB - We report on the performance of two whole genome amplification methods, GenomiPhiTM amplification and modifiedimproved primer extension preamplification (mIPEP), when analysing low template DNA samples. Template as low as 10 pg treated with mIPEP generated more than 1 ng of DNA that could be used in STR typing. Initial templates of 100-10 pg, when treated with mIPEP, generated an increase in alleles compared with control samples. Partial profiles using the AmpF'STRw IdentifilerTM Kit were produced from this suboptimal DNA template, with 70% of the possible alleles (21.7+2.1 in 32 alleles) recorded, using the mIPEP amplified products with an initial template of 100 pg. Allelic imbalance decreased with samples treated with whole genome amplification method (WGA) compared with those without this initial treatment. Further methods for improvement were also analysed including altering the condition of electrokinetic injection, and the successful DNA typing rate was increased to about 80%. This report illustrates the potential use and limitations of WGA for low template samples.
UR - http://www.scopus.com/inward/record.url?scp=84869396340&partnerID=8YFLogxK
U2 - 10.1258/msl.2012.011126
DO - 10.1258/msl.2012.011126
M3 - Article
SN - 0025-8024
VL - 52
SP - 223
EP - 228
JO - Medicine, Science and the Law
JF - Medicine, Science and the Law
IS - 4
ER -