Abstract
We have previously developed the bio-cell chip, on which cells are arrayed and immobilized on a glass slide, using multi-well device. Development of bio-cell chip was originally targeted for the application in fluorescent in situ hybridization and immunohistochemistry. Here, we suggest simple and convenient method of DNA extraction from bio-cell chip, which can handle hundreds of specimens. One uL of cell suspension was loaded on wells of the bio-cell chip and after drying, 2uL of BR-A buffer (GenScript BloodReady™Multiplex PCR System) was dropped onto the wells of bio-cell chip. After incubation for 10 minutes, each mixture was transferred to a microtube with additional buffer to a total volume of 20uL. One uL of mixture was taken and 20uL of mixture for PCR reaction was prepared; 1U BioTherm™DNA Polymerase (Genecraft, GERMANY), 1uL (10pM/ul) of each oligonucleotide primer for target sequence, 2.5mM dNTP, and 1.0uL of DNA mixture. PCR for GST M1 and hGAPDH was carried out using the MJ Research DNA thermal cycler. With this bio-cell chip extraction method, simplification of experimental procedure not only lessened the loss of specimen but also saved the time. Only 10 min of reaction time and a very small volume of samples was required without any further purification step. Amplification of DNA was possible, reflecting safety in DNA stability. Requirement for the diagnostic specimen transport is intricate; strict regulations on infectious materials and preparation of specimen package, cell fixatives such as methanol. Bio-cell chip enables transport of hundreds of samples at a thin plate without transportation media, and also simple extraction of DNA from bio-cell chip with high throughput efficiency, with excellent stability comparable to those with fresh samples.
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