Selective control of primer usage in multiplex one-step reverse transcription PCR

BMC Molecular Biology

Table 1 PCR Primer and hydrolysis probe sequences for the five human total RNA targets.

Target (Length)	NCBI Accession Number	Transcript length (nt)	primer and Probe Information
			primer/probe type	Sequence (5'-3')	5' position	Limit of detection
PBGD (265 bp)	NM_000190.3	1526	Forward primer	GAGTGATTCGCGTGGGTACC	213	n.d.
			Reverse primer	GGCTCCGATGGTGAAGCC	476
			Hydrolysis probe	-	-
ABCA5 (82 bp)	NM_172232.2	8177	Forward primer	GGCTGCTATTCTGACCACTCACTATA	4598	< 7.6E+01 copies
			Reverse primer	TTAACTGCCCAGACACCATGAT	4659
			Hydrolysis probe	6 FAM - CAGAGGCTGTCTGTGATCGAGTAGC - BHQ1	4633
ABCA6 (114 bp)	NM_080284.2	5296	Forward primer	CCATGAGAAATGTCCAGTTTCCT	324	<1.6E+01 copies
			Reverse primer	TGCTGGGTTAAATTAGATATTGGTGTA	412
			Hydrolysis probe	Alexa Fluor^®647-TCCTCAGAATCTGGGAAGGGTAGATAAA-BHQ2	355
ABCA7 (147 bp)	NM_019112.3	6832	Forward primer	TTTCTCTGGGACATGTGTAACTACTTG	4981	<8.9E+01 copies
			Reverse primer	TGTGATCGACCAGCCATACAG	5104
			Hydrolysis probe	HEX C6-NH- CCTTCCAGCAGAGGGCATATGTG - BHQ1	5042
ABCA10 (61 bp)	NM_033450.2	5118	Forward primer	GCGGGTTAAGCTTGTGACAGA	1601	n.d
			Reverse primer	CCCACCCGCAGAACTTGA	1645
			Hydrolysis probe	-	-

To complement the PCR primer and hydrolysis probe sequences, this table also includes the NCBI Accession Number, the amplicon length, the transcript length, the position along the transcript of the primers/probes, and the lowest limit of detection for each of the RNA standards (ABCA5, ABCA6, and ABCA7).

ISSN: 1471-2199