Validation and estimation of parameters for a general probabilistic model of the PCR process.
J Comput Biol
; 14(1): 97-112, 2007.
Article
em En
| MEDLINE
| ID: mdl-17381349
ABSTRACT
Earlier work rigorously derived a general probabilistic model for the PCR process that includes as a special case the Velikanov-Kapral model where all nucleotide reaction rates are the same. In this model, the probability of binding of deoxy-nucleoside triphosphate (dNTP) molecules with template strands is derived from the microscopic chemical kinetics. A recursive solution for the probability function of binding of dNTPs is developed for a single cycle and is used to calculate expected yield for a multicycle PCR. The model is able to reproduce important features of the PCR amplification process quantitatively. With a set of favorable reaction conditions, the amplification of the target sequence is fast enough to rapidly outnumber all side products. Furthermore, the final yield of the target sequence in a multicycle PCR run always approaches an asymptotic limit that is less than one. The amplification process itself is highly sensitive to initial concentrations and the reaction rates of addition to the template strand of each type of dNTP in the solution. This paper extends the earlier Saha model with a physics based model of the dependence of the reaction rates on temperature, and estimates parameters in this new model by nonlinear regression. The calibrated model is validated using RT-PCR data.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Simulação por Computador
/
Reação em Cadeia da Polimerase
/
Modelos Estatísticos
/
Hibridização de Ácido Nucleico
Idioma:
En
Ano de publicação:
2007
Tipo de documento:
Article