Methods of identifyinig dha canola ns-b50027-4
AssigneeNuseed Pty Ltd
InventorsWenxiang Gao, Yonghe Bai, Tina Ngo
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Abstract
The present embodiments provide compositions, methods and primers for detecting DNA of genetically modified canola, specifically of elite event DHA canola NS-B50027-4 and progeny thereof.
Claims17
A qualitative gel-based method of identifying the presence of a DHA canola NS-B50027-4 event or progeny thereof in a biological sample, said method comprising detecting a two event-specific detection method that comprises
- (a) extracting a DNA sample from said biological sample;
- (b) providing a first DNA primer pair combination, wherein said first DNA primer pair combination is selected from (i) nucleic acids as set forth in SEQ ID NO:3 (A05-286F) and SEQ ID NO:4 (A05-286R) or (ii) nucleic acids as set forth in SEQ NO:18 (A05-200F) and SEQ ID NO:19 (A05-200R), respectively;
- (c) providing a second DNA primer pair combination, wherein said second DNA primer pair combination is selected from (i) nucleic acids as set forth in SEQ ID NO:5 (A02-282F) and SEQ ID NO:6 (A02-282R) or (ii) nucleic acids as set forth in SEQ NO:20 (A02-258F) and SEQ ID NO:21 (A02-258R), respectively;
- (d) providing DNA amplification reaction conditions;
- and
- (e) performing two separate DNA amplification reactions with said first (as in step (b)) and second (as in step (c)) DNA primer pair combinations, thereby producing (1) an amplicon pair consisting of a 286-bp amplicon and a 282-bp amplicon, or (2) an amplicon pair consisting of a 200-bp amplicon and a 258-bp amplicon, respectively;
- wherein the presence of said amplicon pair (1) or (2) identifies the presence of the DHA canola NS-B50027-4 event or progeny thereof in the biological sample.
An event-specific quantitative detection method for identifying the presence of a DHA canola NS-B50027-4 event or progeny thereof in a biological sample, said method comprising a two event-specific detection method that comprises:
- (a) extracting a DNA sample from said biological sample;
- (b) providing a first DNA primer pair combination, wherein said first DNA primer pair combination consists of nucleotides as set forth in SEQ ID NO:12 (A05up2F) and SEQ ID NO:13 (A05up2R), respectively;
- (c) providing a second DNA primer pair combination, wherein said second DNA primer pair combination consists of nucleotides as set forth in SEQ ID NO:15 (A02dn2F) and SEQ ID NO:16 (A02dn2R), respectively;
- (d) providing DNA amplification reaction conditions;
- and
- (e) performing two separate DNA amplification reactions with said first and second DNA primer pair combinations, thereby producing a 120-bp amplicon and a 170-bp amplicon, respectively,
- wherein the presence of said 120-bp amplicon and 170-bp amplicon identifies the presence of the DHA canola NS-B50027-4 event or progeny thereof in the biological sample.
A qualitative gel-based method of identifying the presence of a DHA canola NS-B50027-4 event or progeny thereof in a biological sample, said method comprising detecting a two event-specific detection method that comprises
- (a) extracting a DNA sample from said biological sample;
- (b) providing a first DNA primer pair combination, wherein said first DNA primer pair combination is from nucleotides as set forth in SEQ ID NO:28 (A05-216F) and SEQ ID NO:29 (A05-216R);
- (c) providing a second DNA primer pair combination, wherein said second DNA primer pair combination is from nucleotides as set forth in SEQ ID NO:20 (A02-258F) and SEQ ID NO:21 (A02-258R);
- (d) providing DNA amplification reaction conditions;
- and
- (e) performing two separate DNA amplification reactions with said first (as in step (b)) and second (as in step (c)) DNA primer pair combinations, thereby producing an amplicon pair consisting of a 216-bp amplicon and a 258-bp amplicon, respectively;
- wherein the presence of said amplicon pair identifies the presence of the DHA canola NS-B50027-4 event or progeny thereof in the biological sample.
Dependencies are inferred from the claim text.