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Table 2 Primer sequences and main conditions of the PCR methods used

From: Molecular diagnosis of cutaneous leishmaniasis and identification of the causative Leishmania species in Morocco by using three PCR-based assays

   

PCR conditions

Primer name

Primer sequence

Amplification conditions

MgCl2(mM)

dNTP (mM)

Primer (μM)

Unit of Taq DNA Polymerase

DNA (μL)

13A

5′-GTG GGG GAG GGG CGT TCT-3′

94°C for 4 min, 94°C for 1 min, 60°C for 1 min, 72°C for 1 min, 72°C for 10 min (30 cycles)

1.5

0.2

1

1

2.5

13B

5′-ATT TTC CAC CAA CCC CCA GTT-3′

Lmj4

5′-CTA GTT TCC C GC CTC CGA G-3′

94°C for 4 min, 94°C for 1 min, 55°C for 1 min, 72°C for 1 min, 72°C for 10 min (35 cycles)

1.5

0.2

1

1

2.5

Uni21

5′-GGG GTT GGT GTA AAA TAG GCC-3′

LITSR

5′-CTG GAT CAT TTT CCG ATG-3′

94°C for 4 min, 94°C for 40 sec,53°C for 30 sec, 72°C for 1 min, 72°C for 10 min (40 cycles)

2

0.2

0.5

1.5

10

L5.8S

5′-TGA TAC CAC TTA TCG CAC TT 3′

GH20

5′-GAA GAG CCA AGG ACA GGT AC-3′

94°C for 4 min, 94°C for 30 sec, 54.5°C for 1 min, 72°C for 1.3 min, 72°C for 10 min (40 cycles)

2.5

0.2

0.4

1

2

PCO4

5′-CAA CTT CAT CCA CGT TCA CC-3′