Table 1.

Primers used for plasmid construction and qRT-PCR^a

Primer name	Sequence (5′–3′)	Purpose
F2833	ACC GAATTCACGCAGAGTTCACTTTTGTCG	Cloning miR-296 target sequence 2896–2920
R3014	GCA CTCGAG CTTGCAGGTGACATAAATGGC
F2061	ACC GAATTC GTCACTTTTATGTTCACCGGAT	Cloning miR-296 target sequence 2115–2135
R2182	GCA CTCGAG GATTGCAAGCCAAAGTCCC
M2115F	CTTATAGCTTACACACCCCCAGGAGGTCCTTTGC	Generating nucleotide mutations in the miR-296 target sequence 2115–2135
M2115R	GCAAAGGACCTCCTGGGGGTGTGTAAGCTATAAG
M2896F	CTTCAATACATGTTTGTTCCACCCGGAGCCCCCAAAC	Generating nucleotide mutation in the miR-296 target sequence 2896–2920
M2896R	GTTTGGGGGCTCCGGGTGGAACAAACATGTATTGAAG
miR-296 mimics	UGUCCUAACUCCCCCCCGGGA	Overexpression of miR-296
miR-296 inhibitor	UCCCGGGGGGGAGUUAGGACA	Inhibition of miR-296 expression
EVBC-1468F	CTCAGTTGACAGTGTGCCCTC	Generation of nucleotide mutations in the EV71 BrCr TR strain
EVBC-3320R	GATTGCCGTTCGGCTGGTAC
BC2-2896mutF	TCAATACATGTTTGTTCCACCCGGAGCCCCCAAACCAGACTCCAGAG
BC2-2896mutR	CTCTGGAGTCTGGTTTGGGGGCTCCGGGTGGAACAAACATGTATTGA
BC2-2115mutF	TATAGCTTACACACCCCCAGGAGGTCCTTTGCCTAAAGATAGAGCCAC
BC2-2115mutR	GTGGCTCTATCTTTAGGCAAAGGACCTCCTGGGGGTGTGTAAGCTATA
VP3F	AACAAGCTTCAGGTTTCCCCACTGAATTGAAACCTGGC	Cloning VP3 ORF or generating mutation
VP3R	ATT CCCGGG TTA TTGAATAGTGGCCGTTTGCAAA
VP1F	CCAGAATTCAGGGGACAGAGTGGCAGATGTGATTG	Cloning VP1 ORF or generating mutation
VP1R	AACGGATCCTTA GAGCGTAGTGATTGCCGTTCG
HNMT-2896F	CCAATATATGTTTGTGCCACCCGGGGCCCCTAAGCCAGATTCTAGG	Generation of nucleotide mutations in the EV71-HeN luc infectious clone
EVHN4141F	GCGCGTGCTGAAGAACGAGC
HNMT-2896R	CCTAGAATCTGGCTTAGGGGCCCCGGGTGGCACAAACATATATTGG
EVHN6824R	GGTTGACCACTCTAAAGTTGCCC
HNMT-2115F	TCATAGCCTATACACCGCCAGGGGGCCCTCTGCCCAAGGACCGGGCGA
HNMT-2115R	CGCCCGGTCCTTGGGCAGAGGGCCCCCTGGCGGTGTATAGGCTATGA
EvVP1F	GAGAGTTCTATA GGGGACAGT	qRT-PCR detection of EV71 genomic RNA
EvVP1R	AGCTGTGCTATG TGA ATTAGG AA
GAPDH-F	GAAGGTGAAGGTCGGAGTC	qRT-PCR detection of GAPDH
GAPDH-R	GAAGATGGTGATGGGATTTC

^aBold, site mutant nucleotide; underlining, restriction endonuclease cutting sites.