RNA extraction and RT-PCR
The WBCs were harvested in TRIzol® reagent (Thermo Fisher) and total RNA
was isolated according to the manufacturer’s instructions. cDNA was
prepared from 1 µg total RNA, using the SuperScript®VI reverse
transcriptase with random hexamers, following the manufacturer’s
protocol (Thermo Fisher). RT-PCR was done using the primer pair of
BCAP31ex1F 5’-CACGTTGACTGTGGGAAACTC-3’ and BCAP31ex4R
5’-TTCCGTCACATCATCATACTTCC-3’. All RT-PCR products were gel extracted
and sequenced to confirm the normal splicing, intron inclusion, and exon
skipping forms.