Optimization of DNA segments amplification by polymerase chain reaction
Public support
Provider
Academy of Sciences of the Czech Republic
Programme
Scheme of targeted research and development support
Call for proposals
Program podpory cíleného výzkumu a vývoje 2 (SAV02002-S)
Main participants
Ústav molekulární genetiky AV ČR, v. v. i.
Contest type
VS - Public tender
Contract ID
—
Alternative language
Project name in Czech
Optimalizace procesu amplifikace DNA segmentů pomocí polymerázové řetězcové reakce
Annotation in Czech
Polymerázová řetězcová reakce (PCR) je široce využívanou metodou pro amplifikaci vybraných úseků DNA. S cílem zjednodušit fázi přípravy PCR a zvýšit její citlivost a specificitu, budeme studovat úlohu aditiv na stabilitu roztoků, obsahujících pracovní pufr, nukleotidy a DNA polymerázu v optimálních koncentracích (tzv. Master mixy). Aditiva budou zahrnovat inertní barviva, inhibitory proteáz, glycerol a další komponenty zvyšující hustotu PCR reakční směsi a tetramethylammonium oxalát, který zvyšuje specificitu a výtěžnost PCR (Kovářová and Dráber, Nucleic Acid Research, 28: e70-74, 2000). Bude rovněž studován vliv těchto aditiv na tzv. LA ("long and accurate") PCR. Dále budou srovnány 2 metody používané pro zvýšení specificity PCR, tzv. "hot-start". Získané výsledky budou představovat teoretickou basi pro racionální přístupy pro jednoduché a spolehlivé metody amplifikace DNA v PCR a RNA v RT-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
OECD FORD - equivalent branches <br>(according to the <a href="http://www.vyzkum.cz/storage/att/E6EF7938F0E854BAE520AC119FB22E8D/Prevodnik_oboru_Frascati.pdf">converter</a>)
10603 - Genetics and heredity (medical genetics to be 3)<br>10604 - Reproductive biology (medical aspects to be 3)<br>10605 - Developmental biology<br>10608 - Biochemistry and molecular biology<br>10609 - Biochemical research methods<br>20801 - Environmental biotechnology<br>20802 - Bioremediation, diagnostic biotechnologies (DNA chips and biosensing devices) in environmental management<br>20803 - Environmental biotechnology related ethics<br>20901 - Industrial biotechnology<br>20902 - Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation<br>20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials<br>30101 - Human genetics<br>30401 - Health-related biotechnology<br>30402 - Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)<br>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])<br>30404 - Biomaterials (as related to medical implants, devices, sensors)<br>30405 - Medical biotechnology related ethics<br>40401 - Agricultural biotechnology and food biotechnology<br>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<br>40403 - Agricultural biotechnology related ethics
Completed project evaluation
Provider evaluation
U - Uspěl podle zadání (s publikovanými či patentovanými výsledky atd.)
Project results evaluation
New additives for amplification of DNA in coloured reaction mixes for PCR have been found. New monoclonal antibodies specific for Taq DNA polymerase have been prepared. One of these antibodies has been used in a new commercially available product.
Solution timeline
Realization period - beginning
Jan 1, 2002
Realization period - end
Jan 1, 2005
Project status
U - Finished project
Latest support payment
—
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP06-AV0-IB-U/01:1
Data delivery date
Jun 19, 2006
Finance
Total approved costs
2,028 thou. CZK
Public financial support
1,960 thou. CZK
Other public sources
68 thou. CZK
Non public and foreign sources
0 thou. CZK