Optimization and reproducibility of random amplified polymorphic DNA in human.
In this work, we have optimized random amplified polymorphic DNA (RAPD) for the use of human DNA in altering the concentration of the reaction components and the steps of the thermal profile in the polymerase chain reaction. By using two primers in every reaction and 2.5 U Taq DNA polymerase, we found that DNA concentrations between 50 and 500 ng gave reproducible banding patterns. The tested DNA was extracted in seven different ways giving the same amplification results in six of them. We have also observed that reactions consisting of 35 cycles gave sufficient product yields. A slow heating/ramping from the annealing to the extension temperature increased the number of amplified bands and enhanced reproducibility. We conclude that RAPD is a robust and, under the mentioned conditions, reproducible method that could prove very useful for scientists and physicians.
Benter
T
T
Department of Hematology and Oncology, Medical School Hannover, Germany.
Papadopoulos
S
S
Pape
M
M
Manns
M
M
Poliwoda
H
H
D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D009695:Nucleic Acid Renaturation; D012194:RNA-Directed DNA Polymerase; D019105:Random Amplified Polymorphic DNA Technique; D015203:Reproducibility of Results; D019914:Taq Polymerase
D019914:Taq Polymerase; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase