What is transgene copy number?

What is transgene copy number?

Transgene copy number is defined as the number of exogenous DNA insert(s) in the genome. For example, if the exogenous DNA fragment inserts only once at a single locus of the genome, it is a single copy transgenic event. The copy number is closely relevant to another concept, zygosity.

Which technique you would use to detect the copy number of a transgene in genetically modified crops?

real-time quantitative PCR
The method described here uses real-time quantitative PCR to determine the transgene copy number present in the genome of the transformant. Specifically, this method measures the relative transgene copy number by comparing it with an endogenous gene with a known copy number.

How do you calculate copy number variation?

Copy number variation can be detected with various types of tests such as fluorescent in situ hybridization, comparative genomic hybridization and with high-resolution array-based tests based on array comparative genomic hybridization (or aCGH), SNP array technologies and high resolution microarrays that include copy …

What is transgene expression?

A transgene is an artificial gene, manipulated in the molecular biology lab that incorporate all appropriate elements critical for gene expression generally derived from a different species, for example, production of α1-proteinase inhibitor protein in transgenic sheep carrying transgene of human origin.

How do I find my plasmid copy number?

Plasmid copy number was determined by comparing the quantification signal from the plasmid to those from the chromosome. Copy number was then calculated by using a known copy number plasmid as a standard.

How do you test for Transgene?

The two basic methods to detect the presence of a gene/transgene are ELISA and PCR amplification. Both methods have already been described and are robust, although each has advantages and disadvantages. For example, ELISA detects the presence of a gene product (protein) and thus requires an expressing gene.

What are the 3 PCR approaches for GMO identification?

Current analytical methods for the detection, identification, and quantification of transgenic DNA in food and feed are reviewed. These methods include quantitative real-time PCR, multiplex PCR, and multiplex real-time PCR.

How many copies of the eGFP transgene can be detected by qPCR?

A standard curve constructed from serial dilutions of a plasmid containing the EGFP transgene was (i) highly reproducible, (ii) detected as few as two copies, and (iii) was included in each qPCR assay. qPCR analysis of genomic DNA was used to determine transgene copy number in several mouse strains.

How can I determine the transgene copy number of a transformant?

The method described here uses real-time quantitative PCR to determine the transgene copy number present in the genome of the transformant. Specifically, this method measures the relative transgene copy number by comparing it with an endogenous gene with a known copy number.

What is qPCR (quantitative real-time PCR)?

Quantitative real-time PCR (qPCR) is a powerful technique that precisely measures and monitors the amplification of a specific nucleic acid sequence in a DNA sample.

What kind of mice are used for transgene copy number determination?

C57BL/6-Tg (UBC-GFP) 30Scha/J (Ubq-EGFP) mice were also used for transgene copy number determinations. All mice were obtained from Jackson Laboratory (Bar Harbor, ME, USA). See Online Protocol available at www.BioTechniques.com.