Analysis of renin-angiotensin aldosterone system gene polymorphisms in malaysian essential hypertensive and type 2 diabetic subjects

Table 1 Oligonucleotides for amplification and screening for three polymorphisms using PCR-RFLP method

Gene polymorphism	Forward primer (FP) Reverse primer (RP)	PCR cycling conditions	Restriction endonuclease	PCR product (bp)	Restriction fragment size (bp)
A20C	FP-5'-AGA GGT CCC AGC GTG AGT GTC-3' RP-5'-AGC CCA CAG CTC AGT TAC ATC-3'	95°C- 3 m	EcoO109I[10]	265	A-265 C-205,137
		94°C- 1 m
		64°C -1 m
		72°C -1 m
		72°C-5 m
		X 30
MboI	FP-5'-GAG GTT CGA GTC GGC CCC CT-3' RP-5'-TGC CCA AAC ATG GCC ACA CAT-3'	94°C- 5 m	MboI[30]	250	MboI (-) -250
		94°C- 30 s			MboI (+) -171, 79
		68°C- 30 s
		72°C- 30 s
		72°C-5 m
		X 30
Lys173Arg	FP- 5'-AGG CAG CTT CTA CCA GGG CCC CAG TCA CT-3' RP- 5'-CCC CTC CCC TGC AAA TCT CAT CCC TTA-3'	94°C- 3 m	Bsu36I[31]	1286	L-1286, A- 1037, 249
		94°C -45 s
		61°C -45 s
		72°C -1 m
		72°C-10 m
		X 35

PCR cycling conditions are represented as temperature and time [minute (m), seconds (s] of initial denaturation, denaturation, annealing, extension and final extension X number of cycles.