Extraction and Sequencing of Oocyte RNA

RNA from oocytes, blastocysts and EPCs was isolated using the ARCTURUS® PicoPure® RNA Isolation Kit (ThermoFisher Scientific), as per manufacturer's instructions. First strand cDNA synthesis was performed using the qScript Flex cDNA kit (Quantabio, MA, U.S.A), according to manufacturer's protocol. The resultant cDNA was used to amplify target genes by PCR (Primer sequences; Supplementary Table 1). The expression of Ddx4 was confirmed by Sanger sequencing using a previously described protocol [66 (link)].

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Tsai T.S., Johnson J., White Y, & John J.C. (2017). The molecular characterization of porcine egg precursor cells. Oncotarget, 8(38), 63484-63505.

Publication 2017

qScript Flex cDNA kit

Blastocysts Cdna Epcs Genes Isolation Oocytes Primer Synthesis

Corresponding Organization :

Other organizations : Hudson Institute of Medical Research, Monash University, OvaScience (United States)

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Variable analysis

independent variables

RNA isolation method (ARCTURUS® PicoPure® RNA Isolation Kit)
CDNA synthesis method (qScript Flex cDNA kit)

dependent variables

Expression of target genes, including Ddx4

control variables

Manufacturer's instructions for RNA isolation and cDNA synthesis were followed

controls

Sanger sequencing was used to confirm the expression of Ddx4, which was previously described in the literature ([66 (link)])

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