Revised genomic structure of the human ghrelin gene and identification of novel exons, alternative splice variants and natural antisense transcripts

Table 1 Designations and sequences of primers used in RT-PCR

Primer name	Primer sequence (5'-3')	Exon	Ta (°C)	PCR Cycles
V0-F	GAACATTTTTAGTTCCCAAGGAATG	0
V0-R	CTCTCTGGTGTTCAGGGCTC	1	62	40
EX-1-F	AAGATAACACAGCTTTGCACAG	-1
EX-1-R	CTCTGGTGTTCAGGGCTCAG	1	59	40
Sure-F	TTCGTCACTCCGTGAATCAG	N/A
Outer-R	TGGCGTTTGTTCTAAACCAG	0	59 → 57	25 (10 → 15)
Inner-R	GGCTGATGTACATTCCTTGG	0	59	35
Exon-1-Full-F	GTGGATGTTTACTTGCTGGTG	-1/-1* #
Exon-1-Full-R	GTTTGAACATTTATTCGCCTCC	4/4* #	61	35, 40
V-long-ex0-F	GAGGATGATCAGGAGCAGTC	0
V-long-ex0-R	CAGTTTGAACATTTATTCGCCTCC	4	59	40
B2MG-F	TGAATTGCTATGTGTCTGGGT	2
B2MG-R	CCTCCATGATGCTGCTTACAT	3	55	25
cRNA-exon-1-F	CCCAAAGATAACACAGCTTTGCAC	-1
cRNA-exon-1-R	CTAATACGACTCACTATAGGGAGA TGGACCCTGGAGGCCTCT	-1	64	30
CF264800-F	ATGAGCGCTTCTAAACTTAGAG	4/4*
CF264800-R	GCTCCTGTTTCCTAAGATGTC	2*	59	30, 35, 40
ex2*-nested-South-F	GCATTTGCCTCAGCGGT	2*
ex2*-nested-South-R		2*	59	30
5'adapter-out-F	CTGATGGCGATGAATGAACAC	N/A
5'OS-out-R	TCTCTAAGTTTAGAAGCGCTC	4*	59	35
5'adapter-in-F	ATGAATGAACACTGCGTTTGCT	N/A
5'OS-in-R	TCTAAGTTTAGAAGCGCTCATCTG	4*	61	35

T7 promoter sequence in primer cRNA-exon-1-R is underlined. Primers in exon -1 and 4 also span antisense exons (-1* and 4*, respectively) as indicated where appropriate (#).

ISSN: 1471-2164