Gene chip used for identifying pathogenic bacteria in blood and its making method
A technology of gene chips and pathogenic bacteria, applied in the field of gene chips, can solve the problems of missing pathogenic bacteria and poor accuracy, and achieve the effects of accurate diagnosis, avoiding the cultivation stage, and high information content
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[0030] 1. Design of Pathogen-Specific Probes
[0031] The 23S ribosomal RNA gene of bacteria was selected as the target sequence, and species, genus-specific and general sequences were selected from it to design probes for the detection of pathogenic bacteria. The probes are shown in the sequence listing.
[0032] 2. Synthetic probes: Synthesized using a synthesizer from Shanghai Bioengineering Co., Ltd.
[0033] 3. Probe treatment: Since the designed probe is relatively short, it is difficult to solidify on the membrane, so we selectively added a polyT (oligo-deoxythymonucleotide) tail to the 3' end, so that it can be solidified on the nylon membrane superior.
[0034] 4. Preparation of gene chip
[0035] Use the nylon membrane with positive point load produced by Roche, and use the sample spotter to spot samples in a preset order.
[0036] 5. Cross-link the spotted membrane under a long-wave ultraviolet lamp for 5 minutes, so that the probe is firmly fixed on the membran...
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