microRNAs targeting DEAD-box helicases are involved in salinity stress response in rice ( Oryza sativa L.)

BMC Plant Biology

Table 1 Sequences of oligonucleotide primers

Cleavage site mapping^a
OsABP	1^st PCR	5′-ATCAAAATCATATCCCCTCCGC-3′	258 bp
OsABP	Nested PCR	5′-TCCTCCCTCCATACGAAAATTCTG-3′	144 bp
OsDBH	1^st PCR	5′-GAAAAGGGGAACAGTGCTAG-3′	150 bp
OsDBH	Nested PCR	5′-AACCAGTTCAGCTTTGGCTGCT-3′	100 bp
OsDSHCT	1^st PCR	5′-GGCAGGTGGCAGGAAGATTATA-3′	490 bp
OsDSHCT	Nested PCR	5′-ACATCGCCGGTCATCAATCCGACAT-3′	310 bp
QRT-PCR for gene amplification
OsABP	Forward	5′-GGGGGAGGATGGTGAGTAAT-3′	211 bp
OsABP	Reverse	5′-CCACCACCATCATCATCAAA-3′	211 bp
OsDBH	Forward	5′-ACGATGGCACCTCAGAGAAT-3′	177 bp
OsDBH	Reverse	5′-TGGACCAAGTGACAGATGGA-3′	177 bp
OsDSHCT	Forward	5′-GAGGAGGTGGAGAACACGAG-3′	237 bp
OsDSHCT	Reverse	5′-GTAGGCCTTGGCCATCTC-3′	237 bp
α-Tubulin	Forward	5′-GGTGGAGGTGATGATGCTTT-3′	200 bp
α-Tubulin	Reverse	5′-ACCACGGGCAAAGTTGTTAG-3′	200 bp
QRT-PCR for miRNA amplification ^b
osa-MIR414	Forward	5′-TCATCCTCATCATCATCGTCC-3′	100 bp
osa-MIR164e	Forward	5′-GGGAGTTCTGTGATTGGAGAG-3′	100 bp
osa-MIR408	Forward	5′-TGGAGAAGCAGGGCACGTGA-3′	100 bp
U6	Forward	5′-CGATAAAATTGGAACGATACAGA-3′	150 bp
U6	Reverse	5′-ATTTGGACCATTTCTCGATTTGT-3′	150 bp

^a Gene-specific reverse primers used in combination with the FirstChoise RLM-RACE Kit (Ambion, Invitrogen) forward and nested primers.
^b Specific reverse primers for miRNAs, used in combination with the Universal qPCR forward primer provided in the NCode VILO kit (Invitrogen).

ISSN: 1471-2229