FIGURE 6.

Uracil-DNA-glycosylase (UDG)-supplemented reverse transcription–loop-mediated isothermal amplification (RT-LAMP): the system (Kim et al. 2016) removes carryover DNA contamination from one experiment (N) to subsequent ones (N + 1). In the first experiment, uracil is incorporated into contaminants through the use of approximately one-third of the deoxyuridine triphosphate (dUTP): approximately two-thirds of the deoxythymidine triphosphate (dTTP) is used in the amplification reaction—amplicons so derived contain a mixture of T and U bases. In the subsequent (N + 1) experiment, UDG is added to the input sample before amplification. The UDG specifically cleaves uracil-containing contaminants that were inadvertently carried over from the first (N) experiment at room temperature. Upon elevation of the reaction to approximately 65°C, the UDG is heat inactivated, ensuring that only the target RNA (or 100% thymine-containing DNA) target is amplified.