The expression level of the GFRa1 Mm01253716_m1 gene in SSCs and TSCs
was examined by the Fluidigm Biomark system. Glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) Mm99999915-g1 was the reference gene for normalization. SSCs and TSCs were picked
up with a micromanipulator technique, lysed with a solution of lysis buffer that contained
9 μl RT-PreAmp Master Mix (5.0 μl Cells Direct 2× Reaction Mix, Invitrogen, USA), 2.5 μl
0.2× assay pool and 1.3 μl TE buffer, 0.2 μl RT/Taq Superscript III (Invitrogen, USA).
Then, the amount of the amplified product of RNA-targeted copies was examined with TaqMan
real-time PCR on a BioMark Real-Time Quantitative PCR (qPCR) system. Samples were analysed
in two technical repeats. The Ct values were calculated using Excel and GenEx software
(20 -22 (link)).
was examined by the Fluidigm Biomark system. Glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) Mm99999915-g1 was the reference gene for normalization. SSCs and TSCs were picked
up with a micromanipulator technique, lysed with a solution of lysis buffer that contained
9 μl RT-PreAmp Master Mix (5.0 μl Cells Direct 2× Reaction Mix, Invitrogen, USA), 2.5 μl
0.2× assay pool and 1.3 μl TE buffer, 0.2 μl RT/Taq Superscript III (Invitrogen, USA).
Then, the amount of the amplified product of RNA-targeted copies was examined with TaqMan
real-time PCR on a BioMark Real-Time Quantitative PCR (qPCR) system. Samples were analysed
in two technical repeats. The Ct values were calculated using Excel and GenEx software
(20 -22 (link)).
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