Estimating the Copy Number of Transgenes in Transformed Cotton by Real-Time Quantitative PCR

2018 ◽  
pp. 137-157 ◽  
Author(s):  
Chengxin Yi ◽  
Yan Hong
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Real Time Quantitative Pcr

Screening of copy number polymorphisms in human β-defensin genes using modified real-time quantitative PCR

2006 ◽  
Vol 308 (1-2) ◽  
pp. 231-240 ◽  
Author(s):  
QiXing Chen ◽  
Malte Book ◽  
XiangMing Fang ◽  
Andreas Hoeft ◽  
Frank Stuber
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Real Time Quantitative Pcr

scholarly journals Quantification of Mitochondrial DNA Deletion, Depletion, and Overreplication: Application to Diagnosis

2003 ◽  
Vol 49 (8) ◽  
pp. 1309-1317 ◽  
Author(s):  
Béatrice Chabi ◽  
Bénédicte Mousson de Camaret ◽  
Hervé Duborjal ◽  
Jean-Paul Issartel ◽  
Georges Stepien
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Southern Blot ◽  
Quantitative Pcr ◽  
Blot Analysis ◽  
Copy Number ◽  
Southern Blot Analysis ◽  
Mtdna Copy Number ◽  
Mtdna Depletion ◽  
Real Time Quantitative Pcr

Abstract Background: Many mitochondrial pathologies are quantitative disorders related to tissue-specific deletion, depletion, or overreplication of mitochondrial DNA (mtDNA). We developed an assay for the determination of mtDNA copy number by real-time quantitative PCR for the molecular diagnosis of such alterations. Methods: To determine altered mtDNA copy number in muscle from nine patients with single or multiple mtDNA deletions, we generated calibration curves from serial dilutions of cloned mtDNA probes specific to four different mitochondrial genes encoding either ribosomal (16S) or messenger (ND2, ND5, and ATPase6) RNAs, localized in different regions of the mtDNA sequence. This method was compared with quantification of radioactive signals from Southern-blot analysis. We also determined the mitochondrial-to-nuclear DNA ratio in muscle, liver, and cultured fibroblasts from a patient with mtDNA depletion and in liver from two patients with mtDNA overreplication. Results: Both methods quantified 5–76% of deleted mtDNA in muscle, 59–97% of mtDNA depletion in the tissues, and 1.7- to 4.1-fold mtDNA overreplication in liver. The data obtained were concordant, with a linear correlation coefficient (r2) between the two methods of 0.94, and indicated that quantitative PCR has a higher sensitivity than Southern-blot analysis. Conclusions: Real-time quantitative PCR can determine the copy number of either deleted or full-length mtDNA in patients with mitochondrial diseases and has advantages over classic Southern-blot analysis.

Accurate and objective copy number profiling using real-time quantitative PCR

Methods ◽  
2010 ◽  
Vol 50 (4) ◽  
pp. 262-270 ◽  
Author(s):  
Barbara D’haene ◽  
Jo Vandesompele ◽  
Jan Hellemans
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Real Time Quantitative Pcr

scholarly journals SNPs and real-time quantitative PCR method for constitutional allelic copy number determination, the VPREB1 marker case

2011 ◽  
Vol 12 (1) ◽  
Author(s):  
Marcello Frigerio ◽  
Elena Passeri ◽  
Tiziana de Filippis ◽  
Daniela Rusconi ◽  
Rea Valaperta ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Real Time Quantitative Pcr ◽  
Copy Number Determination ◽  
Pcr Method ◽  
Allelic Copy Number

Corrigendum to “Accurate and objective copy number profiling using real-time quantitative PCR” [Methods 50 (2010) 262–270]

Methods ◽  
2011 ◽  
Vol 53 (3) ◽  
pp. 326
Author(s):  
Barbara D’haene ◽  
Jo Vandesompele ◽  
Jan Hellemans
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Real Time Quantitative Pcr

scholarly journals Determination of Cytochrome P450 2D6 (CYP2D6) Gene Copy Number by Real-Time Quantitative PCR

2005 ◽  
Vol 2005 (3) ◽  
pp. 248-253 ◽  
Author(s):  
Laurent Bodin ◽  
Philippe H. Beaune ◽  
Marie-Anne Loriot
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Gene Dosage ◽  
Gene Copy Number ◽  
Rapid Identification ◽  
Gene Copy ◽  
Cyp2d6 Gene ◽  
Real Time Quantitative Pcr ◽  
Gene Copies

Gene dosage by real-time quantitative PCR has proved to be accurate for measuring gene copy number. The aim of this study was to apply this approach to the CYP2D6 gene to allow for rapid identification of poor and ultrarapid metabolizers (0, 1, or more than 2 gene copy number). Using the2−ΔΔCtcalculation method and a duplex reaction, the number of CYP2D6 gene copies was determined. Quantitative PCR was performed on 43 samples previously analyzed by Southern blotting and long PCR including 20 samples with a heterozygous deletion, 11 with normal copy number (2 copies), and 12 samples with duplicated genes. The average ratio ranged from1.02to1.28,1.85to2.21, and2.55to3.30, respectively, for the samples with 1 copy, 2 copies, and 3 copies. This study shows that this method is sensitive enough to detect either a heterozygous gene deletion or duplication.

scholarly journals Quantification of MYCN, DDX1, and NAG Gene Copy Number in Neuroblastoma Using a Real-Time Quantitative PCR Assay

2002 ◽  
Vol 15 (2) ◽  
pp. 159-166 ◽  
Author(s):  
Katleen De Preter ◽  
Frank Speleman ◽  
Valérie Combaret ◽  
John Lunec ◽  
Geneviève Laureys ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Copy Number ◽  
Gene Copy Number ◽  
Gene Copy ◽  
Pcr Assay ◽  
Real Time Quantitative Pcr ◽  
Quantitative Pcr Assay
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