TCR或BCR高通量测序的方法及利用标签序列矫正多重PCR引物偏差的方法

本发明提供了一种对TCR或BCR进行高通量测序的方法，该方法根据TCR或BCR的V区基因特征设计上游引物，根据TCR或BCR的C区或J区基因特征设计下游引物，结合多重PCR技术和高通量测序，获得TCR或BCR的序列，从而分析TCR或BCR的重排信息；采用本发明提供的2个循环的多重PCR技术，相比现有多重PCR的25～30个循环，能大大降低引物扩增偏好引入的测序误差；此外，本发明还提供了一种利用DNA标签序列矫正多重PCR的引物偏差的方法，进一步降低引物扩增偏好引入的测序误差以及高通量测序固有的测序错误。

The invention provides a method for carrying out high-throughput sequencing on a TCR (T cell receptor) or a BCR (B cell receptor). The method is characterized by designing upstream primers according to gene features of a V region of the TCR or the BCR and designing downstream primers according to gene features of a C region or a J region of the TCR or the BCR and obtaining sequences of the of the TCR or the BCR in combination with the multiplex PCR (polymerase chain reaction) technology and high-throughput sequencing, thus analyzing the rearrangement information of the TCR or the BCR. Compared with 25-30 cycles of existing multiplex PCR, two cycles of the multiplex PCR technology provided by the invention can conduce to greatly reducing the sequencing errors caused by primer amplification preference. Besides, the invention also provides a method for correcting multiplex PCR (polymerase chain reaction) primer deviation by utilizing DNA (deoxyribonucleic acid) tag sequences, thus further reducing the sequencing errors caused by primer amplification preference and intrinsic sequencing errors of high-throughput sequencing.