In TGI's data, 730,000 of 1,080,000 human unique consensuses are singleton. The total number of human genes is about 25,000, among which 35~60% contain alternative splicing, so the number of singleton should be about 10,000~15,000. Obviously, most sequences in the singleton are the result of errors. The real singleton sequences perhaps are already included in the tentative consensuses (TC). So we discarded all unique singletons and only count the number of TCs and obtain the result of 328,301. As ASPicDB only analyzed 18,193 genes, while human beings have 25,000 genes. If other genes do not have alternative splicing, then the size of total transcriptome should be 326,552, which is quite close to the result of TGI. It is a pity that only human data in ASPicDB is fairly comprehensive, the data of other species are quite sparse and cannot be used in the same manner.