New generation of preparations for amplifications of DNA fragments by polymerase chain reaction
Project goals
Amplification of DNA fragments by polymerase chain reaction (PCR) is extensively used in basic research as well as for diagnostic and commercial purposes. With increasing numbers of applications of PCRs, the need for stabile formulations of complex PCR mixtures is growing. Our preliminary experiments indicated that lyophilization of complex PCR mixtures (containing DNA polymerases, nucleotides, buffer, anti-polymerase monoclonal antibodies and additives) decreased performance of PCR. To further simplifyPCR set up and to overcome problems with lyophilization we will test the stabilizing effect of additives on rehydratation and dehydratation of complex PCR mixtures. The additives will include sugars, polyethylenglycol, Ficol and others. Furthermore, wewill search for more advantageous properties of mutagenized Taq DNA polymerase. This research could provide a theoretical background for production of a new generation of preparations for amplification of DNA fragments by PCR.
Keywords
polymerase chain reactionTaq DNA polymeraselyophilizationmutagenesis
Public support
Provider
Academy of Sciences of the Czech Republic
Programme
The supprot of targeted research projects (National proramme of research)
Call for proposals
Podpora projektů cíleného výzkumu 1 (SAV02005-CI)
Main participants
—
Contest type
VS - Public tender
Contract ID
1QS500520551
Alternative language
Project name in Czech
Nová generace přípravků pro amplifikaci DNA fragmentů pomocí polymerázové řetězcové reakce
Annotation in Czech
Amplifikace DNA fragmentů polymerázovou řetězcovou reakcí (PCR) je extenzivně využívaná jak v základním výzkumu tak v diagnostice a pro komerční účely. S narůstajícím počtem aplikací PCR se zvyšují požadavky na formulaci stabilních přípravků pro PCR. Naše předběžné výsledky ukázaly, že lyofilizace kompletních reakčních směsí pro PCR (obsahujících DNA polymerázy, nukleotidy, pufr, anti-polymerázové monoklonální protilátky a aditiva) snižuje účinnost PCR. Abychom dále zjednodušili přípravu rekčních směsípro PCR a řešili problémy s lyofilizací budeme testovat stabilizační efekt aditiv na dehydrataci a rehydrataci komplexních směsí pro PCR. Aditiva budou zahrnovat cukry, polyethylenglykol, Ficol a další. Dále budeme hledat nové výhodnější formy mutagenizované Taq DNA polymerázy. Tento výzkum bude představovat teoretickou bazi pro produkci nové generace přípravků pro amplifikaci DNA fragmentů v PCR.
Scientific branches
R&D category
NV - Nonindustrial research (Applied research excluded Industrial research)
CEP classification - main branch
EB - Genetics and molecular biology
CEP - secondary branch
EI - Biotechnology and bionics
CEP - another secondary branch
CE - Biochemistry
10603 - Genetics and heredity (medical genetics to be 3)
10604 - Reproductive biology (medical aspects to be 3)
10605 - Developmental biology
10608 - Biochemistry and molecular biology
10609 - Biochemical research methods
20801 - Environmental biotechnology
20802 - Bioremediation, diagnostic biotechnologies (DNA chips and biosensing devices) in environmental management
20803 - Environmental biotechnology related ethics
20901 - Industrial biotechnology
20902 - Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation
20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials
30101 - Human genetics
30401 - Health-related biotechnology
30402 - Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
30403 - Technologies involving identifying the functioning of DNA, proteins and enzymes and how they influence the onset of disease and maintenance of well-being (gene-based diagnostics and therapeutic interventions [pharmacogenomics, gene-based therapeutics])
30404 - Biomaterials (as related to medical implants, devices, sensors)
30405 - Medical biotechnology related ethics
40401 - Agricultural biotechnology and food biotechnology
40402 - GM technology (crops and livestock), livestock cloning, marker assisted selection, diagnostics (DNA chips and biosensing devices for the early/accurate detection of diseases) biomass feedstock production technologies, biopharming
40403 - Agricultural biotechnology related ethics
Completed project evaluation
Provider evaluation
U - Uspěl podle zadání (s publikovanými či patentovanými výsledky atd.)
Project results evaluation
This project led to formulation of new reaction mixes for quantitative polymerase chain reaction (qPCR). These reaction mixes possess all constant components of qPCR and allow amplification even of long genomic DNA fragments.
Solution timeline
Realization period - beginning
Jul 1, 2005
Realization period - end
Dec 31, 2009
Project status
U - Finished project
Latest support payment
Mar 11, 2009
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP10-AV0-1Q-U/01:1
Data delivery date
Apr 15, 2010
Finance
Total approved costs
4,077 thou. CZK
Public financial support
3,735 thou. CZK
Other public sources
0 thou. CZK
Non public and foreign sources
342 thou. CZK
Recognised costs
4 077 CZK thou.
Public support
3 735 CZK thou.
0%
Provider
Academy of Sciences of the Czech Republic
CEP
EB - Genetics and molecular biology
Solution period
01. 07. 2005 - 31. 12. 2009