SINGLE TUBE MULTIPLEX ASSAY FOR DETECTION OF ADULTERANTS IN BASMATI RICE SAMPLES

• US 20110086764A1
• Filed: 07/23/2010
• Published: 04/14/2011
• Est. Priority Date: 04/08/2002
• Status: Abandoned Application

• Current Assignee: Centre For Dna Fingerprinting and Diagnostics
• Original Assignee: Centre For Dna Fingerprinting and Diagnostics
• Inventors: ARCHAK, SUNIL, NAGARAJU, JAVAREGOWDA
• Application Number:
  US12/842,746
• Publication Number:
  US 20110086764A1
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: C12Q 1/6888 for detection or identification of organisms
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present invention provides a single tube multiplex assay for distinguishing basmati from non-basmati rice varieties, and thereby identifying the adulteration of basmati rice varieties. The present invention further provides a method for quantifying adulteration in basmati rice varieties. The present invention also provides a kit for performing a multiplex assay for distinguishing basmati from non-basmati rice varieties. The kit may comprise a primer directed to an SSR loci, appropriate reagents for PCR, and optionally, a package insert for conducting the assay.

Claims

1-3. -3. (canceled)

4. 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 μ

l;

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.

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

a) amplifying 10 ng/10 μ

l 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%.