A simple and rapid gene amplification from Arabidopsis leaves using AnyDirect system.
Polymerase chain reaction (PCR) is a powerful technique in molecular biology and is widely used in various fields. By amplifying DNA fragments, PCR has facilitated gene cloning procedures, as well as molecular genotyping. However, the extraction of DNA from samples often acts as a limiting step of these reactions. In particular, the extraction of PCR-compatible genomic DNA from higher plants requires complicated processes and tedious work because plant cells have rigid cell walls and contain various endogenous PCR inhibitors, including polyphenolic compounds. We recently developed a novel solution, referred to as AnyDirect, which can amplify target DNA fragments directly from whole blood without the need for DNA extraction. Here, we developed a simple lysis system that could produce an appropriate template for direct PCR with AnyDirect PCR buffer, making possible the direct amplification of DNA fragments from plant leaves. Thus, our experimental procedure provides a simple, convenient, non-hazardous, inexpensive, and rapid process for the amplification of DNA from plant tissue.
Yang
Young Geun
YG
Department of Biochemistry, College of Science, Yonsei University, Seoul 120-749, Korea.
Kim
Jong Yeol
JY
Soh
Moon-Soo
MS
Kim
Doo-Sik
DS
D017360:Arabidopsis / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D005784:Gene Amplification; D018515:Plant Leaves / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D015203:Reproducibility of Results
D018744:DNA, Plant