Single tube multiplex assay for detection of adulterants in basmati rice samples

A single tube multiplex assay for distinguishing basmati from non-basmati rice varieties and thereby quantifying adulteration, said assay comprising steps of:

• a) amplifying the RM171 and RM72 locus of DNA from basmati and non-basmati rice varieties using forwarding primers having SEQ ID NO.:
• 18 and SEQ ID NO.:
• 21, respectively and reverse primers having SEQ ID NO.:
• 28 and SEQ ID NO.:
• 31, respectively, and a DNA template in the amount of 10 ng/10 μl;
• b) determining a ratio of amplified products of basmati and non-basmati rice varieties;
• c) constructing a standard curve on the basis of ratio of quantity of amplified products of the alleles of adulterant the non-basmati rice and the basmati rice varieties against a progressive proportion of the adulteration;
• and
• d) quantifying the adulteration in the basmati rice variety on the basis of peak area of the alleles corresponding to basmati and that of the adulterant non-basmati rice varieties, wherein the non-basmati rice is quantified with an accuracy of at least ±3% adulteration.

A method of quantifying adulteration in basmati rice varieties, said method comprising steps of:

• a) amplifying 10 ng/10 μl of DNA from basmati and non-basmati rice varieties;
• b) determining a ratio of amplified products of the basmati and the non-basmati rice varieties;
• c) constructing a standard curve on the basis of quantity of the amplified products;
• and
• d) quantifying the adulteration in the basmati variety, with an accuracy of atleast ±3%.