A New Nested Allele-Specific Multiplex Polymerase Chain Reaction Method for Haplotyping of VKORC1 Gene to Predict Warfarin Sensitivity

Table 2

Primers used in PCR1 and PCR2.

	Sequence (5′ to 3′)	Location	Length (bp)	Product size (bp)	𝑇𝑚 (°C)
Primers for PCR1
381 & 861
Com (F)	GCCCAGGAGTTAGAGGCAACATAAC	257–281	25	1060	66.2
Com (R)	CAGCTTTCTCTGATCTCCTGGTGTG	1316–1292	25		66.2
5808
Com (F)	ATTCTGGAGTCTGGGATCGGTGTG	5546–5569	24	398	66.3
Com (R)	ACCCCAGAATCTCCAGCTCCCTG	5943–5921	23		68.1
9041
Com (F)	CAGCTCCTGGCATCTAGGTAGTGC	8604–8627	24	853	68.0
Com (R)	CTTCCAGGTGTGTGCTCAGCCTTC	9456–9433	24		68.0
Primers for PCR2
381
381 & 861 Com (R)	CAGCTTTCTCTGATCTCCTGGTGTG	1316–1292	25		66.2
WT (F)	AGCACTTTAGGAAGCCAAGGAGGGC	357–381	25	960	67.9
Mut (F)	AGCACTTTAGGAAGCCAAGGAGGGT	357–381	25	960	66.2
861
381 & 861 Com (R)	CAGCTTTCTCTGATCTCCTGGTGTG	1316–1292	25		66.2
WT (F)	AAACTCCTGACCTCAGGTGATCCAC	837–861	25	480	66.2
Mut (F)	AAACTCCTGACCTCAGGTGATCCAA	837–861	25	480	64.6
5808
Com (F)	ATTCTGGAGTCTGGGATCGGTGTG	5546–5569	24		66.3
WT (R)	CGCCAACACCCCCCTTCA	5825–5808	18	280	71.7
Mut (R)	CGCCAACACCCCCCTTCC	5825–5808	18	280	70.8
9041
Com (R)	CTTCCAGGTGTGTGCTCAGCCTTC	9456–9433	24		68.0
WT (F)	CCTCCTCCTGCCATACCCG	9023–9041	19	434	66.6
Mut (F)	CCTCCTCCTGCCATACCCA	9023–9041	19	434	64.5