| Title |
Genetiškai modifikuotų augalų transgeninių intarpų molekulinis apibūdinimas kuriant, optimizuojant ir taikant polimerazine grandinine reakcija pagrįstus metodus / |
| Translation of Title |
Development, optimization and application of PCR-based methods for molecular characterization of transgenic inserts in genetically modified plants. |
| Authors |
Spalinskas, Rapolas |
| Full Text |
|
| Pages |
27 |
| Keywords [eng] |
GMO ; LT-RADE ; LT-PCR ; genome walking ; carbon nanotubes |
| Abstract [eng] |
One of the fastest developing fields of biotechnology in the past three decades is that of genetically modified (GM) plants for industrial applications, as well as food and feed. The European Union (EU) has probably the most rigorous regulations for GMOs in the world and as such, the decisions made by the EU political institutions on regulatory and marketing actions heavily depend on scientific evidence. This doctoral dissertation deals with the molecular biology laboratory methods based on the most popular to date method polymerase chain reaction (PCR) used in the molecular characterization of the genetically modified organisms (GMOs). When studying the molecular structure of the GMO, two main interests of the nucleotide sequence are: (1) the actual sequence of the transgenic insert, and (2) the plant genomic DNA surrounding the synthetic DNA. Two distinct techniques of the Long Template PCR and Genome Walking were analyzed on the GMO DNA matrices. The LT-PCR was applied to obtain single uninterrupted fragment containing full transgenic insert of the selected GM events. The analysis of LT-PCR was continued with the application of a novel PCR additive called carbon nanotubes (CNTs) to investigate the impact of this nanomaterial on the specificity, efficiency and the product yield. The Genome Walking is a technique to obtain unknown DNA fragments adjacent to the known DNA. This doctoral dissertation seeks to explain in detail the development and optimization of a method Long Template Rapid Amplification of Genomic DNA Ends (LT-RADE) and its application on various GMO DNA matrices. |
| Type |
Summaries of doctoral thesis |
| Language |
Lithuanian |
| Publication date |
2014 |
