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. 2022 Nov 26;2:100105. doi: 10.1016/j.crpvbd.2022.100105

Table 1.

PCR conditions which differed between the published protocol by Sulaiman et al. (2003) and the modified protocol.

Primer name	Primer sequence (5′-3′)	Product length (bp)	Buffer^a	No. of cycles	Annealing T (°C)	Total reaction volume (μl)	Template DNA per reaction (μl)	dNTP (μM)^a	MgCl₂ (mM)^a	Taq (U) per reaction	Primer (nM)^a
AL3543 (Outer forward)	AAATIATGCCTGCTCGTCG	605	1x	35	50.0	100	0.25–2.00	200 each	3	5	200
AL3546 (Outer reverse)	CAAACCTTITCCGCAAACC	605	1x	35	50.0	100	0.25–2.00	200 each	3	5	200
AL3544 (Inner forward)	CCCTTCATCGGIGGTAACTT	530	1x	35	50.0	100	2.50	200 each	3	5	200
AL3545 (Inner reverse)	GTGGCCACCACICCCGTGCC	530	1x	35	50.0	100	2.50	200 each	3	5	200
AL3543Mod (Outer forward)^b	AAATYATGCCTGCTCGTCG	605	10x	40	54.2	20	3.00	50 each	15 (incorporated into buffer)	0.5	500
AL3546Mod (Outer reverse)^b	TGGCCACCACRCCCGTGCC	605	10x	40	54.2	20	3.00	50 each	15 (incorporated into buffer)	0.5	2000
AL3544Mod (Inner forward)^b	CAAACCTTYTCYGCAAACC	531	10x	40	57.8	20	3 at 1:1000 dilution of first round PCR product	50 each	15 (incorporated into buffer)	0.5	500
AL3545Mod (Inner reverse)^b	CCCTTCATCGGYGGTAACTT	531	10x	40	57.8	20	3 at 1:1000 dilution of first round PCR product	50 each	15 (incorporated into buffer)	0.5	500

Abbreviation: T, temperature.

^a

Concentration.

^b

New primers.