+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Determination of transgene copy number by real-time quantitative PCR

Determination of transgene copy number by real-time quantitative PCR

Methods in Molecular Biology 526: 129-134

Efficient methods to characterize transgenic plants are important to quickly understand the state of the transformant. Determining transgene copy number is an important step in transformant characterization and can differentiate between complex and simple transformation events. This knowledge can be extremely useful when determining what future experiments and uses the transgenic lines can be utilized for. 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. This method is a quick alternative to the Southern blot, a method that is commonly used to determine gene copy number, and is effective when screening large numbers of transformants.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 052537791

Download citation: RISBibTeXText

PMID: 19378009

DOI: 10.1007/978-1-59745-494-0_11

Related references

Standard addition quantitative real-time PCR (SAQPCR): a novel approach for determination of transgene copy number avoiding PCR efficiency estimation. Plos one 8(1): E53489, 2013

Improvement of peanut Arachis hypogaea L transformation efficiency and determination of transgene copy number by relative quantitative real-time PCR. In Vitro Cellular & Developmental Biology - Plant 49(3): 266-275, 2013

Estimating transgene copy number by real-time quantitative PCR in transgenic maize. Plant Biology (Rockville) : 182, 2001

High-throughput assessment of transgene copy number in sugarcane using real-time quantitative PCR. Plant Cell Reports 31(1): 167-177, 2012

Quantitative real-time PCR assay for determining transgene copy number in transformed plants. Biotechniques 31(1): 132-4, 136-40, 2001

Estimation of transgene copy number in transformed citrus plants by quantitative multiplex real-time PCR. Biotechnology Progress 24(6): 1241-1248, 2009

Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum). Analytical Biochemistry 375(1): 150-152, 2008

Real-time quantitative PCR-based system for determining transgene copy number in transgenic animals. Biotechniques 37(4): 610-613, 2004

Quantitative real-time PCR as a screening tool for estimating transgene copy number in WHISKERS derived transgenic maize. Plant cell reports 20(10): 948-954, 2002

Transgene copy number comparison in recombinant mammalian cell lines: critical reflection of quantitative real-time PCR evaluation. Cytotechnology 65(5): 811-818, 2013

Determination of cytochrome P450 2D6 (CYP2D6) gene copy number by real-time quantitative PCR. Journal of Biomedicine & Biotechnology 2005(3): 248-253, 2005

Using real-time PCR to determine transgene copy number in wheat. Plant Molecular Biology Reporter 22(2): 179-188, 2004

Quantitative real-time polymerase chain reaction for determination of plasmid copy number in bacteria. Journal of Microbiological Methods 65(2): 258-267, 2006

Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes. Microbial Cell Factories 7: 6, 2008

Real-Time PCR for the Detection of Precise Transgene Copy Number in Wheat. Methods in Molecular Biology 1679: 251-257, 2017