Total RNA (2 μg)
was reversetranscribed
into
cDNA
using
an iScript Advanced
cDNA
Synthesis kit
for RT-qPCR
(Bio-Rad Laboratories, Hercules, CA, USA),
according to
the manufacturer's instructions.
Real-time PCR
was performed using
SsoAdvanced SYBR Green Supermix (Bio-Rad).
The CDC10 mRNA
expression levels in the LM
were determined with
the MiniOpticon
Real-Time PCR Detection System (Bio-Rad),
using the CDC10 mRNAspecific
primers (F: 5′-TCTGAAGCTGAGCTCCAA-3′ and R: 5′-AAATTGGT
CAAACGGATCCA-3′).