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.
Ma
Hao
H
Department of Biology, West Virginia University, Morgantown, WV 26506, USA. hao.ma@mail.wvu.edu
Difazio
Stephen
S
D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D032107:Populus / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*