Methods for Sequencing Immune Repertoires
A technology of immune group library and immunoglobulin, applied in the field of quantitative nucleic acid analysis
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[0022] The present invention provides a method of sequencing the improvement of the immunoholic library. The previous method of determining the immunohistocket library, for example, those methods based on multiple PCRs are described in WO 2011 / 140433 and WO 2012 / 083069. Multiple PCRs have many limitations that make it particularly useless to accurately determine the immunoholic library. (see figure 2 These limitations include capturing preferences and amplification preferences caused by PCR. Multiple PCR techniques of the sequencing immunohistochemistry are designed to initiate primers in all frame areas of known V gene fragments. When the mutation occurs at the initiation position, the capture preferences occur, and the mutated gene amplification is insufficient. PCR preferences are generated due to the relative amounts of PCR replicates having the same sequences. Therefore, PCR preferences can result in significant cloning or diversity lacking. In general, the observed group lib...
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