Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids

Table 1

List of primers used in this study for the generation of capture probes and quantitative real-time reverse transcriptase PCR.

Primer name	Sequence (5′ → 3′)	Length (bp)	PCR condition
18S R	GATCCTCTAGAACAGCAGCCG	85	See Section 2.2
28S R	ATCCTTCGATGTCGGCTCTTC	100
D*	NH2-GTTTCCCAGTAGGTCTCNNNNNNNN		See Section 2.3
T3	NH2-GTTTCCCAGTAGGTCTCNNNNNNNN		See Section 2.2
BR18S-F^#	AGGAATTCCCAGTAAGTGCG	102	30 cycles of 94°C,15′′;
BR18S-R^#	GCCTCACTAAACCATCCAA		60°C, 1′
28SF4006	CGCCGGTGAAATACCACTAC	200	35 cycles of 95°C, 15′′;
28SR4205	CTGAGCTCGCCTTAGGACAC		55°C, 20′′; 72°C, 30′′
tdcA-F^@	CGGTGGTGGAAGTCTCATTT	173	35 cycles of 95°C, 10′′;
tdcA-R^@	ACCAATCGCAAAATCCAGTC		54°C, 20′′; 72°C, 20′′

Note: *published primer from Wang et al. 2002 [32]. ^#Published primer pair from Grace et al. 2003 [33]. ^@Published primer pair from Lin et al. 2010 [34]. All other primer pairs are novel to this study. The length indicates amplicon size.