Authentic single cell RNA-Seq by measuring the transcriptome of an individual cell twice. Mus musculus

We developed a novel single-cell RNA-Seq technique that can directly measure the technical noises for each individual gene by splitting a single cell into two equal halves and measuring their transcriptomes separately (Measure a single cell twice by RNA-Seq, MAST-Seq).The MAST-Seq protocol was developed from the single cell RNA-Seq technique we previously developed (Tang et al. 2010). We first lysated the cell to release the RNA from the cell, then reverse transcripted the mRNA into cDNA, added polyA to the 3' end of cDNA, synthesized the second strand of cDNA used designed primer, and at last PCR amplified the double strand cDNA to get enough amount of DNA to construct the cDNA libraries. Overall design: 10 single mouse ES cells, 4 half of single mouse ES cells, 4 quarter of single mouse ES cells, 8 eighth of single mouse ES cells, 4 quarter of 2 single mouse ES cells, 4 quarter of 2 single MEF cells,5 single mouse inner cell mass cells, 5 mouse trophectoderm cells, 4 half of single zygotes, 12 half of single 2-cell embryo blastomeres, 24 half of single 4-cell embryo blastomeres, 8 eighth of 8-cell embryo and 64 half of single 8-cell embryo blastomeres, in total 156 samples.