Whole blood immune repertoire detection method based on high-throughput sequencing technology
An immunological library and detection method technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as inconvenience and directness, and achieve the effect of low cost and good sequencing time.
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[0022] The whole blood immune repertoire detection method based on high-throughput sequencing technology provided by the present invention comprises the following steps:
[0023] (1) RNA is directly extracted from whole blood samples;
[0024] (2) Generate single-stranded cDNA from RNA by reverse transcription, connect the oligonucleotide linker sequence to the 3' end of the single-stranded cDNA, and use the general primer and the special targeting primer P-VDJ with the sequence GACCTCGGGTGGGAACAC as the special primer for PCR reaction The single-stranded cDNA connected with the oligonucleotide linker sequence is used as the PCR template to carry out PCR reaction, and the immune receptor genome is amplified;
[0025] (3) High-throughput sequencing.
[0026] The method provided by the present invention will be further described below in conjunction with examples.
[0027] Using previously collected whole blood samples from two malignant tumor patients and two benign patients,...
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