‘Touchdown’ PCR to circumvent spurious priming during gene amplification

Errors induced during PCR amplification

The Polymerase Chain Reaction (PCR) is one of the most widely used techniques in modern molecular biology to amplify a single or few copies of a piece of DNA sequences. The PCR products are used as inputs of many other applications such as diagnosis of diseases. Therefore the accuracy of the further analysis depends on the quality of the PCR outputs. In order to study the amplification process ...

Single-step PCR optimization using touchdown and stepdown PCR programming.

Method To Detect Only Live Bacteria during PCR Amplification.

Ethidium monoazide (EMA) is a DNA cross-linking agent and eukaryotic topoisomerase II poison. We previously reported that the treatment of EMA with visible light irradiation (EMA + Light) directly cleaved chromosomal DNA of Escherichia coli (T. Soejima, K. Iida, T. Qin, H. Taniai, M. Seki, A. Takade, and S. Yoshida, Microbiol. Immunol. 51:763-775, 2007). Herein, we report that EMA + Light rando...

Gene amplification during myogenic differentiation

Gene amplifications are mostly an attribute of tumor cells and drug resistant cells. Recently, we provided evidence for gene amplifications during differentiation of human and mouse neural progenitor cells. Here, we report gene amplifications in differentiating mouse myoblasts (C2C12 cells) covering a period of 7 days including pre-fusion, fusion and post-fusion stages. After differentiation in...

Type-specific Amplification of 5S rRNA from Panax ginseng Cultivars Using Touchdown (TD) PCR and Direct Sequencing

Generally, the direct sequencing through PCR is faster, easier, cheaper, and more practical than clone sequencing. Frequently, standard PCR amplification is usually interpreted by mispriming internal or external regions of the target template. Normally, DNA fragments were eluted from the gel using Gel extraction kit and subjected to direct sequencing or cloning sequencing. Cloning sequencing ha...