Evaluation of recombinase polymerase amplification for detection of begomoviruses by plant diagnostic clinics

Table 3 Effect of type of template and cleaning method on the usefulness of amplicons generated by recombinase polymerase amplification (RPA) of Tomato yellow leaf curl virus using primer pair TYL828F/TYL834R in three downstream applications

Template	Amplicon treatment	TA Cloning efficiency (%)^a	Restriction endonuclease digestion	Direct sequencing
Purified DNA	No treatment	0	yes	nt^b
	denatured 65 °C	0	no	nt
	denatured 65 °C then centrifuged	80	yes	2/2
	PCR purification column	100	yes	1/2
Crude extraction	No treatment	0	yes	nt
	denatured 65 °C	0	no	nt
	denatured 65 °C then centrifuged	0	yes	2/2
	PCR purification column	100	yes	2/2

^a Cloning efficiency is defined as the number of colonies with the expected insert in ten randomly selected white colonies
^b nt = Not tested

ISSN: 1743-422X