Table 1 List of primers used in this study
A) Primers used for human miRNAs precursors amplification | ||||
Genes | Forward Primer (5′-3′) | Reveres Primer (5′-3′) | ||
miR-125b | GTCGACTGTGAAGGAAAGGAGC | TCTAGAACTTTAACAGAAATCCAGG | ||
miR-637 | GTCGACACCCCAGGGATGATG | TCTAGA AGGCGGAGCACAGAAG | ||
miR-802 | GTCGACCAGTTTGTTCCAGTGCC | TCTAGATAACACACACAGCCCCAG | ||
Scramble | GTCGACCCGCTTGTTCGTTGGTAACTACATTCAAGAGATGTAGTTACC | GTCTAGAAAAAAGCTTGTTCGTTGGTAACTACATCTCTTGAATGTAGTTACC | ||
B) Oligonucleotide sequences used for construction of pSICHECK2.MCS | ||||
Name | (5′to3′) | |||
OligoF | TCGACCGGAGCTCATACTAGTACACCGGTAACTCGAGTAGC | |||
OligoR | GGCCGCTACTCGAGTTACCGGTGTACTAGTATGAGCTCCGG | |||
C) Primers used for site-directed mutagenesis PCR | ||||
Mutated site | Forward Primer (5′-3′) | Reveres Primer (5′-3′) | ||
miR-125b | F2:GCCCGCTCTCTCTTGGTTGGACCTGC F3:AAAGCAAAGGGCGTACCCCCACATTCTCTfC | R2:GTCCAACCAAGGGCCTGAGAGAGCG R3:GAATGTGGGGGTACGCCCTTTGCTTTCC | ||
miR-637 | F2:CTTGGCAGCCCCTCACCCCCCTTC | R2:GCCTCATAAAGAGGGACACACAGAAG | ||
miR-802 | F2:CCTCAGACACCGTAAGCTTGCAGGCCTCAG F3:GCAATAATCTTTGAGATGCGCGGCTGTTC | R2:TTCACTCCAGCTATGCAC R3:GAACAGCCGCGCATCTCAAAGATTATTGCTTTGTG | ||
Common primer | F1:GTCGACAGCGCCCCAGCCTGCA | R1:CTCGAGAACGTTTTCAAATAATTTATTAGG | ||
D) Primers used for gene expression analysis by Q-PCR | ||||
Genes | Forward Primer (5′-3′) | Reveres Primer (5′-3′) | TM (°C) | Accession No |
MSI1 | TTGGCAGACTACGCAGGAAG | TGGTCCATGAAAGTGACGAAGC | 59 | NM_002442 |
Gapdh | CCACTCCTCCACCTTTGACG | CCACCACCCTGTTGCTGTAG | 60 | NM_001357943 |
E-cadherin | CTGCTGCTCTTGCTGTTTCTTC | CTTCTCCGCCTCCTTCTTCATC | 60 | NM_001317185 |
EpCAM | CCATGTGCTGGTGTGTGAAC | CCTTCTGAAGTGGTCCGC | 60 | NM_002354 |
CD24 | ATGTGGCAAGGAAAAACAGG | TTGGCATCCATCATCTAGTC | 60 | NM_001359084 |
Vimentin | AAACTTAGGGGCGCTCTTGT | TGAGGGCTCCTAGCGGTTTA | 59 | NM_003380 |
Snail | CCAGAGTTTACCTTCCAGCA | GATGAGCATTGGCAGCGA | 60 | NM_005985 |