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. 2015 May 15;10(5):e0127475. doi: 10.1371/journal.pone.0127475

Table 1. Sequences of primers used for PCR, sequencing and Real-Time PCR, followed by sequences of a TaqMan probe, shRNAs, and CRNDEP epitopes used in the present study.

Primer name Sequence [5' → 3'] 1 or [NH2 → COOH]
L388-SacI PCR primer CCTGAGCTCCTATGAGAGGATCGCATCAC
L388-SmaIv2 PCR primer GGTCCCGGGCAATTAAGCTTTAGTCTATAAAC
L388BglII PCR primer GGTAGATCTCTATGAGAGGATCGCATCAC
L388SmaI PCR primer GGTCCCGGGCATTAAGCTTTAGTCTATAAAC
RedL388F PCR primer AACCACAACGAGGACTAC
RedL388R PCR primer GGACAAACCACAACTAGAATG
L388wewF forward sequencing primer CTGTACTGGCTATTGGAAGG
L388wewR reverse sequencing primer TAAACCACTCGAGCACTTTG
U6prom-F sequencing primer GACTATCATATGCTTACCGT
SV40rev sequencing primer CATACTTCTGCCTGCTGGG
LOCrtF—Real-Time PCR forward primer (universal) AATTCATCCCAAGGCTGGTC
LOCrt35R —Real-Time PCR reverse primer (specific to the CRNDEP-coding transcript, FJ466686) TTCCAGTGGCATCCTCCTTA
LOCrt4wR—Real-Time PCR reverse primer (specific to non-CRNDEP-coding transcripts, e.g., FJ466685) GCACTCACAATGAGTCATCTG
CRNDE-specific TaqMan probe (universal) CCTTCCAATAGCCAGTACAGTAGCTCC
SH1 sense strand 2 GATCGGGAGCTACTGTACTGGCTATTGGAAGGAGTCAAGAGCTCCTTCCAATAGCCAGTACAGTAGCTCCTTTTTTGA
SH1 complementary strand 2 AGCTTCAAAAAAGGAGCTACTGTACTGGCTATTGGAAGGAGCTCTTGACTCCTTCCAATAGCCAGTACAGTAGCTCCC
SH2 sense strand 2 GATCGAGAAGAAGGTTAAGCTGTATTTGATTGCCTCAAGAGGGCAATCAAATACAGCTTAACCTTCTTCTTTTTTTGA
SH2 complementary strand 2 AGCTTCAAAAAAAGAAGAAGGTTAAGCTGTATTTGATTGCCCTCTTGAGGCAATCAAATACAGCTTAACCTTCTTCTC
SH3 sense strand 2 GATCGGAAGATAAGGAGGATGCCACTGGAAATGTTCAAGAGACATTTCCAGTGGCATCCTCCTTATCTTCTTTTTTGA
SH3 complementary strand 2 AGCTTCAAAAAAGAAGATAAGGAGGATGCCACTGGAAATGTCTCTTGAACATTTCCAGTGGCATCCTCCTTATCTTCC
SH SCR (scrambled, negative control) sense strand 2 GATCGGGAGCAATATCGTGGATGAAACGGTGAAATCAAGAGTTTCACCGTTTCATCCACGATATTGCTCCTTTTTTGA
SH SCR (scrambled, negative control) complementary strand 2 AGCTTCAAAAAAGGAGCAATATCGTGGATGAAACGGTGAAACTCTTGATTTCACCGTTTCATCCACGATATTGCTCCC
CRNDEP Epitope 1 EIHPKAGLQSLQ
CRNDEP Epitope 2 DSEDKEDATGNVEMK
CRNDEP Epitope 3 EWFKYVLGIPVYRL

1) Restriction enzyme recognition sites inserted in primer overhangs were underlined.

2) shRNA-coding sequences: sense regions were marked with a wavy line, whereas antisense regions were indicated with a double wavy line; sticky ends, specific to BamHI/HindIII digestion, were double-underlined.