Nested LAMP (PCR followed by LAMP). LAMP. isoPCR. FIP. F3. F1c. F2. LB. FP. Target DNA. 3’. F3c. F2c. F1c. B1. LBc. B2. LF. 5’. Supplemental Fig. 1
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Nested LAMP (PCR followed by LAMP) LAMP isoPCR FIP F3 F1c F2 LB FP Target DNA 3’ F3c F2c F1c B1 LBc B2 LF 5’ Supplemental Fig. 1 General location of primers in relation to the target DNA. The isoPCR method performed using FIP and BIP primers in a first-stage preamplification PCR, the product of which is detected in distinct isothermal reactions using FIP, BIP, LF and LB primers. Multiplexing is possible by including multiple FIP/BIP primer sets in the first-stage preamplification prior to splitting the product and detecting the specific targets in distinct isothermal reactions. LAMP reactions are performed using conventional F3, FIP, LF, LB, BIP and B3 primer design. Nested LAMP is performed using a first-stage PCR with FP, BP primers followed by a LAMP reaction. Nested qPCR is performed using a first-stage PCR with FP and BP primers followed by a second-stage qPCR reaction with F3 and B3 primers. BP FPc B3 5’ 3’ F3 F2 F1 B1c B2c LFc LB BPc FP B3c LF B2 B1c B3 BP BIP