scholarly journals Real-Time PCR for the Detection of Precise Transgene Copy Number in Wheat

2017 ◽  
pp. 251-257
Author(s):  
Angelica Giancaspro ◽  
Agata Gadaleta ◽  
Antonio Blanco
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Transgene Copy Number

Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum)

2008 ◽  
Vol 375 (1) ◽  
pp. 150-152 ◽  
Author(s):  
Cheng Xin Yi ◽  
Jun Zhang ◽  
Ka Man Chan ◽  
Xiao Kun Liu ◽  
Yan Hong
Keyword(s):  
Gossypium Hirsutum ◽  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Transgene Copy Number

scholarly journals Real-time PCR to determine transgene copy number and to quantitate the biolocalization of adoptively transferred cells from EGFP-transgenic mice

BioTechniques ◽  
2008 ◽  
Vol 45 (3) ◽  
pp. 247-258 ◽  
Author(s):  
Molishree U. Joshi ◽  
H. Keith Pittman ◽  
Carl E. Haisch ◽  
Kathryn M. Verbanac
Keyword(s):  
Transgenic Mice ◽  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Transgene Copy Number

scholarly journals Real-time PCR for the detection of precise transgene copy number in durum wheat

2011 ◽  
Vol 16 (4) ◽  
Author(s):  
Agata Gadaleta ◽  
Angelica Giancaspro ◽  
Maria Cardone ◽  
Antonio Blanco
Keyword(s):  
Durum Wheat ◽  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Gene Copy Number ◽  
Correlation Coefficients ◽  
Gene Copy ◽  
Transgene Copy Number ◽  
Locus Number ◽  
Wheat Lines

AbstractRecent results obtained in various crops indicate that real-time PCR could be a powerful tool for the detection and characterization of transgene locus structures. The determination of transgenic locus number through real-time PCR overcomes the problems linked to phenotypic segregation analysis (i.e. lack of detectable expression even when the transgenes are present) and can analyse hundreds of samples in a day, making it an efficient method for estimating gene copy number. Despite these advantages, many authors speak of “estimating” copy number by real-time PCR, and this is because the detection of a precise number of transgene depends on how well real-time PCR performs.This study was conducted to determine transgene copy number in transgenic wheat lines and to investigate potential variability in sensitivity and resolution of real-time chemistry by TaqMan probes. We have applied real-time PCR to a set of four transgenic durum wheat lines previously obtained. A total of 24 experiments (three experiments for two genes in each transgenic line) were conducted and standard curves were obtained from serial dilutions of the plasmids containing the genes of interest. The correlation coefficients ranged from 0.95 to 0.97. By using TaqMan quantitative real-time PCR we were able to detect 1 to 41 copies of transgenes per haploid genome in the DNA of homozygous T4 transformants. Although a slight variability was observed among PCR experiments, in our study we found real-time PCR to be a fast, sensitive and reliable method for the detection of transgene copy number in durum wheat, and a useful adjunct to Southern blot and FISH analyses to detect the presence of transgenic DNA in plant material.

scholarly journals Quantitative Real-Time PCR Assay for Determining Transgene Copy Number in Transformed Plants

BioTechniques ◽  
2001 ◽  
Vol 31 (1) ◽  
pp. 132-140 ◽  
Author(s):  
David J. Ingham ◽  
Sandra Beer ◽  
Stephanie Money ◽  
Geneviève Hansen
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Transgene Copy Number ◽  
Transformed Plants

scholarly journals Transgene copy number comparison in recombinant mammalian cell lines: critical reflection of quantitative real-time PCR evaluation

2013 ◽  
Vol 65 (5) ◽  
pp. 811-818 ◽  
Author(s):  
Wolfgang Sommeregger ◽  
Bernhard Prewein ◽  
David Reinhart ◽  
Alexander Mader ◽  
Renate Kunert
Keyword(s):  
Real Time ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Real Time Pcr ◽  
Copy Number ◽  
Critical Reflection ◽  
Quantitative Real Time Pcr ◽  
Transgene Copy Number ◽  
Mammalian Cell Lines ◽  
Number Comparison

Using real-time PCR to determine transgene copy number in wheat

2004 ◽  
Vol 22 (2) ◽  
pp. 179-188 ◽  
Author(s):  
Zhiwu Li ◽  
Jennifer L. Hansen ◽  
Ying Liu ◽  
Robert S. Zemetra ◽  
Philip H. Berger
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Transgene Copy Number

Estimation of transgene copy number in transformed citrus plants by quantitative multiplex real-time PCR

2008 ◽  
Vol 24 (6) ◽  
pp. 1241-1248 ◽  
Author(s):  
Ahmad A. Omar ◽  
Marty G. H. Dekkers ◽  
James H. Graham ◽  
Jude W. Grosser
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Transgene Copy Number
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