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用家族特异性免疫球蛋白可变区基因引物两两组合分别对轻、重链进行ＲＴＰＣＲ扩增．在不同退火温度下得到了全部轻链及大部分重链（１３／１６）可变区基因．当先用免疫球蛋白信号肽序列５′端引物与未扩出基因３′端引物组合进行一次ＰＣＲ，再以该产物为模板进行二次ＰＣＲ，则获得了未扩出三条重链的ＰＣＲ产物．这表明通过ＰＣＲ反应条件的改变及程序调整，可增加所获得可变区基因的种类及数量，从而增加抗体库的多样性
Using family-specific immunoglobulin variable region gene primers pairwise combinations of light and heavy chain RT-PCR amplification. All light chains and most heavy chain (13/16) variable region genes were obtained at different annealing temperatures. When the first immunoglobulin signal peptide 5’-end primer was used in combination with the 3’-end primer of the unexpanded gene for PCR, and then the product was used as a template for the second PCR, a PCR product. This indicates that by changing the PCR reaction conditions and adjusting the program, the types and amounts of the obtained variable region genes can be increased to increase the diversity of the antibody library