An amplification primer, kit and application for detecting mutations in genes causing familial retinitis pigmentosa
A technology for retinal pigment and amplification primers, which is applied in the field of amplification primers for detection of mutations in genes causing familial retinitis pigmentosa. problem, to achieve good auxiliary diagnosis value, improve timeliness, and wide coverage
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[0042] Preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings.
[0043] Reagents used:
[0044] Ion Ampliseq Library kit 2.0, Ion PGM Template OT2 200 kit v3, Ion PGM Sequencing 200 kit v2, Ion xpress Barcorde Adaptor33-48 kit, Ion 316 chip kit v2 BC.
[0045] 1. Extract the whole genome DNA of the sample to be tested
[0046] Using OMEGA "Genomic DNA Extraction Kit" to extract peripheral blood DNA. The total volume of extracted DNA is about 100μl, the concentration is 20-100ng / μl; A260 / 280 is between 1.8-2.0. use 2.0 Fluorescence Quantitative Instrument for accurate quantification of genomic DNA.
[0047] 2. PCR amplification of the target region of genomic DNA
[0048] The amount of DNA required for the amplification reaction of each primer set (primer pool 1 and primer pool 2) was 10 ng (2 primer sets in total). The concentration of primers in each primer pool was 400 nM.
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