An efficient method for purification of PCR products for sequencing.
A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1%-0.2% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and labor efficient, provides high quality sequences at a low cost, and bypasses problems with impure PCR products. This technique has been used for single nucleotide polymorphism (SNP) discovery in Populus angustifolia trees.
Department of Biology, West Virginia University, Morgantown, WV 26506, USA. hao.ma@mail.wvu.edu
Hao
H
Ma
Stephen
S
Difazio
D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D032107:Populus / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*