Method of preparing cell lysate
AssigneesHitachi Chemical Company, Ltd., Hitachi Chemical Research Center, Inc.
InventorMasato Mitsuhashi
Life of this patent
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Abstract
The entire process of reverse transcription-polymerase chain reaction (RT-PCR) is simplified by using oligonucleotide-immobilized microplates made of, e.g., polypropylene, to which oligonucleotides are securely immobilized and which can be subjected to thermal cycles of PCR. RT-PCR is preferably conducted in solid-phase. Capturing of mRNA and RT-PCR can be conducted in the same plates. The cDNA synthesized from the mRNA captured on the microplates can be used more than once. Further, in combination with the microplates, a filter plate is used for the preparation of cell lysates wherein target cells are placed on the filter plate, and a lysis buffer is passed through the cell layer on the filter to transfer cell lysate directly to the microplate via well-to-well communication.
Claims3
A method of preparing cell lysate from target cells, comprising the steps of:
- transferring target cells to a filter plate provided with a membrance having a pore size such that the target cells are trapped, but cytosolic mRNA present in the cells passes therethrought, wherein the target cells are places on the membrance;
- and
- passing a lysis buffer through the cell layer on the membrane to obtain a cell lysate of the target cell.
Dependencies are inferred from the claim text.