GSM7124508: run_13_seq_1_PA-04B_S26; Mus musculus; RNA-Seq
To generate the dataset, we used the optimized sci-RNA-seq3 protocol, adjusting the volume and type of lysis buffer to the size of the embryos. Briefly, frozen embryos were pulverized on dry ice and cells were lysed with a phosphate-based, hypotonic lysis buffer containing magnesium chloride, Igepal, diethyl pyrocarbonate (DEPC) as an RNase inhibitor, and either sucrose or bovine serum albumin (BSA). Lysate is passed over a 20um filter, and the nuclei-containing flow-through is fixed with a mixture of methanol and dithiobis (succinimidyl propionate) (DSP). Nuclei are rehydrated and washed in a sucrose/PBS/triton/magnesium chloride buffer (SPBSTM), then counted and distributed into 96-well plates for reverse transcription with indexed oligo-dT primers.