NDLI: A non-PCR-based Invader$^{®}$ assay quantitatively detects single-copy genes in complex plant genomes

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A non-PCR-based Invader$^{®}$ assay quantitatively detects single-copy genes in complex plant genomes

Content Provider	Springer Nature Link
Author	Gupta, Manju Nirunsuksiri, Wilas Schulenberg, Greg Hartl, Thomas vak, Stephen Bryan, Jill Vapdorp, Nathan Bing, James Thompson, Steve
Copyright Year	2007
Abstract	An accurate determination of gene copy number is critical to the success of a molecular breeding program involving both transgenic and non-transgenic plants. In this paper, we have described the application of a non-PCR-based technology, Invader$^{®*}$, for determination of gene copy number and zygosity in plants. A biplex assay format detected both a target gene and an endogenous reference gene simultaneously from the genomic DNA. The ratio between the signals of the two genes in relation to known copy number standards of the same target gene allowed copy number determination. The linear range of the Invader assay was 1–4 copies per genome, but it can be accurate over a larger copy number range depending on the assay conditions. This technique was utilized for screening plants carrying low transgene copy numbers from a large number of events generated by plant transformation, and shown to produce results comparable to that of Southern blots. We have also utilized this technique to screen thousands of field-grown plants for zygosity determinations and obtained data that was over 98% accurate, thus proving that this assay can be used to improve the efficiency of a breeding program. Overall, the Invader assays proved to be reproducible, specific, applicable to any gene sequence and amenable to high-throughput screening.
Starting Page	173
Ending Page	181
Page Count	9
File Format	PDF
ISSN	13803743
Journal	Molecular Breeding
Volume Number	21
Issue Number	2
e-ISSN	15729788
Language	English
Publisher	Springer Netherlands
Publisher Date	2007-07-26
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Cry1F Gene copy number Invader assay Non-PCR-based single-copy gene detection Trait introgression Zygosity assay Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science Molecular Biology Biotechnology Agronomy and Crop Science

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Use of real-time PCR for determining copy number and zygosity in transgenic plants

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Copy Number of Bean Mitochondrial Genes Estimated by Real-time PCR does not Correlate with the Number of Gene Loci and Transcript Levels

Zygosity identification in transgenic cotton (Gossypium hirsutum) by real-time quantitative PCR

Mapping of the Ogura fertility restorer gene Rfo and development of Rfo allele-specific markers in canola (Brassica napus L.)

Easy and rapid method of zygosity determination in transgenic mice by SYBR$^{®}$ Green real-time quantitative PCR with a simple data analysis

A non-PCR-based Invader$^{®}$ assay quantitatively detects single-copy genes in complex plant genomes

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A non-PCR-based Invader$^{®}$ assay quantitatively detects single-copy genes in complex plant genomes