Method and apparatus for inspecting gel chip
A chip detection and gel technology, applied in biochemical equipment and methods, biological testing, material inspection products, etc., can solve the problems of unpaired probes or impurity contamination, non-charged impurities are difficult to remove, and chip sensitivity is not high , to improve signal detection sensitivity, facilitate adjustment of cleaning power, and reduce chip background
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Embodiment 1
[0018] Example 1: Using acrylamide gel chips to simultaneously immobilize PCR products of 1000 individuals to type the same SNP site
[0019] The Y (C / T) in the 5'GCTTTCTTGTTTGTTTTCCTCCTTTACCAY(C / T)CCAGAAATCCATTTGAGTCTGCTCCTTGT-3' sequence is a gene locus related to the mutation of the hyperlipidin gene (Homo sapiens dystrobrevin binding protein 1, DTNBP1). Through a pair of primers (the reverse primer has acrylamide modification), amplify a gene sequence of 1000 people containing this site by PCR, and directly mix 5-10ul of PCR products or purified products with acrylamide (acrylamide monomer and methylene acrylamide monomer), ammonium persulfate, glycerol and water to form a prepolymer, and transfer it to a 384-well plate, and spot 1000 samples on the same glass slide as a microchip by a chip spotter. array. Then the chip was baked at 80°C for 30 seconds and then taken out. The chip was placed in 0.1M NaOH solution and treated with ultrasonic waves for 10 seconds to denatur...
Embodiment 2
[0020] Example 2: Rapid processing of gel chips in gene sequencing by this method
[0021] One universal PCR reverse primer PR was modified with acrylamide, while the other forward primer PF was not modified. Mix the above reverse primer with acrylamide (acrylamide monomer and methylene acrylamide monomer), ammonium persulfate, glycerol and water to form a prepolymer, and spot it on the chip. Then place the chip in a vessel containing TEMED activator for decompression and vacuum filtration. Since TEMED volatilizes to the surface of the gel, ammonium persulfate is excited to release free radicals and the prepolymer is gelled, thereby preparing a compound containing amplification. Primers for gel microarray chips. After breaking a larger genomic DNA with ultrasonic waves, the same universal linker (complementary to the reverse primer immobilized in the gel) was connected to the two ends of each fragment respectively, and then used PCR solution ( Containing Taq enzyme, buffer, ...
Embodiment 3
[0022] Example 3: Using the device and method to process the chip on the agarose protein chip in the process of antigen antibody detection for 650 people
[0023] First melt the agarose, spread it evenly on the surface of the glass slide, treat it with sodium periodate after solidification, and clean it. Chlamydia trachomatis protein was precipitated after centrifugation from 650 human urine samples, then diluted with PBS buffer, and spotted on the chip with a spotter, and left to stand at 37°C for 30 minutes, and the chip was placed in an ultrasonic generator Sonicate for 5s and rinse with PBS. Spread the antibody with streptavidin evenly on the surface of the chip and react at 37°C for 30 minutes, sonicate for 5 s and rinse with PBS, and then mix the fluorescent Cy3 labeled with biotin with the antigen 28 of the labeled avidin antibody. Cultivate for 1 hour. Finally, the chip was placed in an ultrasonic generator and ultrasonicated for 5 seconds, rinsed with PBS, and final...
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