| 主权项 |
1. A process for the preparation of a biological sample in a closed system comprising a lysis chamber fluidly connected to a first PCR chamber containing a restrained magnet inside the first PCR chamber, a second PCR chamber fluidly connected to the first PCR chamber, and a reagent chamber containing dried PCR reagents and fluidly connected to the first PCR chamber, the process comprising:
a) digesting the biological sample in a lysis buffer in the lysis chamber; b) binding nucleic acids of the digested biological sample to magnetic particles in the lysis chamber; c) separating the magnetic particles having the bound nucleic acids from the lysis buffer; d) transferring the magnetic particles having the bound nucleic acids to the first PCR chamber; e) introducing water to the reagent chamber to dissolve the dried PCR reagents and transporting the dissolved reagents to the first PCR chamber; f) amplifying non-specifically the nucleic acids bound to the magnetic particles in a first PCR of a nested PCR in the first PCR chamber to produce unbound nucleic acid strands; g) separating the unbound nucleic acid strands in a supernatant from the nucleic acids bound to the magnetic particles with the restrained magnet disposed in the internal space of the first PCR chamber; h) transferring the supernatant to the second PCR chamber comprising PCR reagents; and i) amplifying specifically the unbound nucleic acid strands in a second PCR of the nested PCR in the second PCR chamber, wherein the lysis chamber, first PCR chamber, second PCR chamber and reagent chamber along with the fluidic connection between the lysis chamber and first PCR chamber, the fluidic connection between the first PCR chamber and reagent chamber, and the fluidic connection between the first PCR chamber and second PCR chamber, form a closed fluidic system; and wherein the restrained magnet is confined within the first PCR chamber. |
