title: GSM3730985: CT-4 (Blastocyst-190); Homo sapiens; RNA-Seq
library_construction_protocol: Human embryos at different stages were treated with Tyrode's acidic solution to remove the zona pellucida then seperated by 0.25% typsin digestion. Single blastomeres were washed twice with 1xPBS containing 0.1% BSA and transferred to 0.2ul of lysis buffer (0.5% NP40, 1U RNase inhibitor) by mouth pipetting. Whole-transcript amplification for single blastomeres was done using the Quartz-seq method. Briefly, cDNA was first generated by reverse transcription with SuperScript III (Life technologies, 18080-044); After the digestion of the redundant RT-primer with exonuclease I (TAKARA, 2650A), first strand of cDNA was poly-A tailed with terminal transferase (Roche Applied Science, 3333574); After synthesis of the second strand of cDNA with tagging primer and Mighty DNA polymerase (TAKARA, R071A), the whole transcriptome was amplified usingsuspension PCR. PCR products were purified with Qiagen MinElute PCR purification kit (QIAGEN, 28004) and Library construction was performed following Illumina's instruction. Libraries were sequenced on the Illumina Hiseq 2000 platform.
attributes.tag: GEO Accession
attributes.value: GSM3730985