Integrated analysis system and dna chip of real-time polymerase chain reaction and integrated analysis method using the same
A technology of chain reaction and polymerase, applied in chemical instruments and methods, biochemical equipment and methods, analytical materials, etc., can solve the problems of time inconvenience, a lot of labor, and difficult molecular diagnosis, so as to improve reliability and prevent sample The effect of evaporation
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Embodiment 1
[0080] [Example 1] Preparation of primers and probes
[0081] (1) Preparation of primers for distinguishing Mycobacterium tuberculosis / Nontuberculosis Mycobacteria from each other, probes for real-time PCR, and primers and probes for DNA chips for diagnosing antibiotic resistance
[0082] The oligonucleotide primers, probes for real-time PCR, and DNA chip probes used to distinguish Mycobacterium tuberculosis and diagnose resistance to rifampicin (rpoB) and isoniazid (inhA, katG) are shown in Table 1 ( tuberculosis), Table 2 (probes for real-time PCR for differentiating Mycobacterium tuberculosis), Table 3 (primers for diagnosing antibiotic resistant immobilized DNA chip probes).
[0083] The probes shown in Table 1 and Table 2 were used to quantitatively analyze the reaction products of Mycobacterium tuberculosis amplified by real-time PCR, FAM was used as a fluorescent material, and TAMRA was used as a quenching material. In order to hybridize the amplified reaction produ...
Embodiment 2
[0105] [Example 2] Attaching the probe to the supporter
[0106] Each probe for quantitative and qualitative analysis prepared in Example 1 was diluted to 100 pmol respectively, and then transferred to a 96-well microplate. The spotting solution was added thereto, and mixed again to 50 pmol. A microarray (USA, Cartesian Technologies, PLXSYS7500SQXL microarray spotter) was used to attach the probes to carriers such as glass slides, membranes, and the like.
[0107] To remove probes not attached to the surface of the carrier, the carrier thus obtained was washed with a 0.2% sodium dodecyl sulfate (SDS) solution at room temperature. The support thus obtained was washed at room temperature with sodium borohydride solution and then with boiling distilled water. The carrier thus obtained was washed with a 0.2% SDS solution and distilled water at room temperature, and then the surface of the carrier was completely dried using a centrifugal separator, thereby completing the preparat...
Embodiment 3
[0108] [Example 3] Preparation of biological material detection device
[0109] Fix the biochip prepared in Example 2 to the bottom of the rod to prepare a biomaterial detection device.
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