Skip to main content

Table 1 Parameters of the RT-LAMP reaction

From: One-step detection of Bean pod mottle virusin soybean seeds by the reverse-transcription loop-mediated isothermal amplification

Component (concentration) (supplier)

Volume ( μL) per 10 μL reaction

Autoclaved PCR-grade MilliQ™water

2.8

10 × ThermoPol reaction buffer (containing 2 mM MgCl2) (New England Biolabs)

1

*25 mM MgCl2 (4 mM final) (TaKaRa)

1.6

10 mM dNTP’s (0.8 mM final) (TaKaRa)

0.8

FIP 20 μM (0.8 μM final) (Invitrogen)

0.4

BIP 20 μM (0.8 μM final) (Invitrogen)

0.4

F3 10 μM (0.2 μM final) (Invitrogen)

0.2

B3 10 μM (0.2 μM final) (Invitrogen)

0.2

5 M Betain (0.8 M final) (Sigma)

1.6

Bst DNA polymerase 8 U/μL (0.32 U/μL final) (New England Biolabs)

0.4

AMV reverse transcriptase 10 U/μL (0.05 U/μL final) (Promega)

0.05

DNA/RNA template or Crude RNA extract

0.6

  1. *Total [MgCl2] is 6 mM, as the reaction buffer contains an addtional 2 mM.