Optimization of different parameters of polymerase chain reaction for Arabidopsis thaliana At4g20020 gene

摘要

Optimization of different parameters of polymerase chain reaction (PCR) for Arabidopsis thaliana At4g20020 gene was conducted. Isolated DNA was reverse transcribed using gene specific primers (Forward primer: 5 -GATGGCTATGATATCTCACCGTCTCC-3 Reverse primer: 5-GTTGTTCCAGTCTCGATTCCTCTGAT-3'). Variable bands were found in gel image in respect to different annealing temperature, concentration of template DNA, MgCl_2 and Taq DNA polymerase. The 56 ℃ was found to provide clear DNA band which suggest maximum amplification of sample DNA. In addition to it, 10 ng of template DNA, 3mM MgCl_2 and 1U/20μ1 concentration of Taq DNA polymerase were found to be promising for successful PCR of At4g20020 gene in Arabidopsis thaliana. This approach provides effective tools for any downstream application of PCR product with At4g20020 gene.