NDLI: Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants

Please wait, while we are loading the content...

A Bioinformatics Approach to the Identification of Variants Associated with Type 1 and Type 2 Diabetes Mellitus that Reside in Functionally Validated miRNAs Binding Sites

Fetuin-A is Associated to Serum Calcium and AHSG T256S Genotype but Not to Coronary Artery Calcification

Bioinformatics Analysis of NBS-LRR Encoding Resistance Genes in Setaria italica

Inferring Phylogenetic Relationships of Indian Citron (Citrus medica L.) based on rbcL and matK Sequences of Chloroplast DNA

The Intra-familial Relationships of Pentaphylacaceae s.l. as Revealed by DNA Sequence Analysis

Evaluation of hepatic arterial anatomy by multidetector computed tomographic angiography in living donor liver transplantation

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants

The Evaluation of Serum Copeptin Levels and Some Commonly Seen Thrombophilic Mutation Prevalence in Acute Pulmonary Embolism

SNP Discovery by GBS in Olive and the Construction of a High-Density Genetic Linkage Map

Cyclooxygenase 2 (rs2745557) Polymorphism and the Susceptibility to Benign Prostate Hyperplasia and Prostate Cancer in Egyptians

Biochemical Genetics : Volume 54, Issue 2, April 2016

Biochemical Genetics : Volume 54, Issue 1, February 2016

Biochemical Genetics : Volume 53

Biochemical Genetics : Volume 52

Biochemical Genetics : Volume 51

Biochemical Genetics : Volume 50

Biochemical Genetics : Volume 49

Biochemical Genetics : Volume 48

Biochemical Genetics : Volume 47

Biochemical Genetics : Volume 46

Biochemical Genetics : Volume 45

Biochemical Genetics : Volume 44

Biochemical Genetics : Volume 43

Biochemical Genetics : Volume 42

Biochemical Genetics : Volume 41

Biochemical Genetics : Volume 40

Biochemical Genetics : Volume 39

Biochemical Genetics : Volume 38

Biochemical Genetics : Volume 37

Biochemical Genetics : Volume 36

Biochemical Genetics : Volume 35

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants

Content Provider	Springer Nature Link
Author	Stefa, Biricolti Patrizia, Bogani Matteo, Cerboneschi Massimo, Gori
Copyright Year	2016
Abstract	One of the major unanswered questions with respect to the commercial use of genetic transformation in woody plants is the stability of the transgene expression over several decades within the same individual. Gene expression is strongly affected by the copy number which has been integrated into the plant genome and by the local DNA features close to the integration sites. Because woody plants cannot be subjected to selfing or backcrossing to modify the transgenic allelic structure without affecting the valuable traits of the cultivar, molecular characterization of the transformation event is therefore crucial. After assessing the transgene copy number of a set of apple transgenic clones with Southern blotting, we describe two alternative methods: the first is based on inverse PCR (i-PCR) and the second on the quantitative PCR (q-PCR). The methods produced comparable results with the exception of the data regarding a high copy number clone, but while the q-PCR-based system is rapid and easily adaptable to high throughput systems, the i-PCR-based method can provide information regarding the transformation event and the characteristics of the sequences flanking the transgenic construct.
Starting Page	291
Ending Page	305
Page Count	15
File Format	PDF
ISSN	00062928
Journal	Biochemical Genetics
Volume Number	54
Issue Number	3
e-ISSN	15734927
Language	English
Publisher	Springer US
Publisher Date	2016-02-19
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Southern blot Inverse PCR Quantitative PCR Transgene copy number Transgenic fruit trees Human Genetics Biochemistry Zoology Medical Microbiology
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Biochemistry Molecular Biology Ecology, Evolution, Behavior and Systematics

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants.

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

High-Throughput and Accurate Determination of Transgene Copy Number and Zygosity in Transgenic Maize: From DNA Extraction to Data Analysis

Determination of Transgene Copy Number in Stably Transfected Mammalian Cells by PCR-Capillary Electrophoresis Assay

High-throughput transgene copy number estimation by competitive PCR

High-throughput assessment of transgene copy number in sugarcane using real-time quantitative PCR

Integration and inheritance of transgenes in crop plants and trees

Relevance of BAC transgene copy number in mice: transgene copy number variation across multiple transgenic lines and correlations with transgene integrity and expression

A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants

Similar Documents

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants.

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

High-Throughput and Accurate Determination of Transgene Copy Number and Zygosity in Transgenic Maize: From DNA Extraction to Data Analysis

Determination of Transgene Copy Number in Stably Transfected Mammalian Cells by PCR-Capillary Electrophoresis Assay

High-throughput transgene copy number estimation by competitive PCR

High-throughput assessment of transgene copy number in sugarcane using real-time quantitative PCR

Integration and inheritance of transgenes in crop plants and trees

Relevance of BAC transgene copy number in mice: transgene copy number variation across multiple transgenic lines and correlations with transgene integrity and expression

A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot

Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants