In vivo quantification of formulated and chemically modified small interfering RNA by heating-in-Triton quantitative reverse transcription polymerase chain reaction (HIT qRT-PCR)

Table 3 Oligonucleotides and TaqMan probes for the stem-loop real time PCR assays.

Assay	Primer/probe	Sequence
AD1596 AD1661	Stem-loop RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAGGAT
	Forward primer	GCCCGCGTAAGACTTGAGATGATC
	Reverse primer	GTGCAGGGTCCGAGGT
	TaqMan probe	(6-FAM) CTGGATACGACAAGGAT (MGB)
AD6490	Stem-loop RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAGGGAA
	Forward primer	CCCGCTTGGATCAAATATAAGATT
	Reverse primer	GTGCAGGGTCCGAGGT
	TaqMan probe	(6-FAM) CTGGATACGACAGGGAA (MGB)
AD1955	Stem-loop RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAACTTA
	Forward primer	TCGTGTTCGAAGTACTCAGCGTAA
	Reverse primer	GTGCAGGGTCCGAGGT
	TaqMan probe	(6-FAM) CTGGATACGACAACTTA (MGB)
AD1661 (sense)	Stem-loop RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAGTAA
	Forward primer	GCCCGCGGATCATCTCAAGTCTTA
	Reverse primer	GTGCAGGGTCCGAGGT
	TaqMan probe	(6-FAM) CTGGATACGACAAGTAA (MGB)

All assays designed to detect the antisense strands of the indicated duplexes, except for the indicated assay designed for sense strand detection. All sequences are written in the 5'-3' direction.
6-FAM = 6-carboxyfluorescein; MGB = minor groove binder; RT = reverse transcription.