Method for simultaneously sequencing multi-sample CDR3 (complementary determining region 3) receptor library with high flux
A high-throughput, multi-sample technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high sample preparation cost, cumbersome operation, heavy workload, etc., and achieve high-throughput sequencing cost reduction , the effect of extensive application value
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[0013] The technical solution of the present invention will be described in detail below in conjunction with the examples.
[0014] Take high-throughput sequencing analysis of the human TCR β chain CDR3 receptor library as an example:
[0015] 1. The human TCR β chain V gene family contains 45 functional gene families and 7 ORFs, and 36 upstream primers are designed according to the homology of 52 genes (see Table 1, 45 or 52 or other similar primers can also be designed source family combination).
[0016] 2. Primer design of multiple base compositions of the C and J genes of the human TCR β chain:
[0017] 1) According to the C gene of human TCR β chain (TRBC1 / TRBC2, highly homologous, see Table 2), multiple downstream primers suitable for pairing with upstream primers can be designed at the front end of the TRBC gene, such as:
[0018] (1) TRBC ttctgatggc tcaaacac;
[0019] (2) TRBC gaagcagag atctcccac;
[0020] (3) TRBC acctgaacaa ggtgttccca;
[0021] (4) TRBC ccacacc...
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